SEQLAB – The best thing that can happen to a DNA.

Transcrição

SEQLAB – The best thing
that can happen to a DNA.
www.seqlab.de
CONTENTS
SEQLAB
Who we are
What we do
GENOMICS
Our sequencing service
Your order (info and tips)
Your data
Your advantages
Our purifying systems
Our favourite thermocycler
PROTEOMICS
Our polyclonal antibodies
Our protein identification
and sequencing
Our peptide synthesis
SERVICE
Price list
Order forms
3
4
5
6
8
8
9
12
13
14
15
16
19
SEQLAB 03
WHO WE ARE
SEQLAB is the DNA service in Germany. Our speciality is DNA sequencing
performed by our highly effective service as seen in our tenders. SEQLAB,
founded in 1996 possesses extensive experience in this field. No DNA is
too sparse, no PCR-product too contaminated with secondary bands and
no band shadow too difficult for us to clone.
Five scientists and ten laboratory assistants are dedicated to fulfilling the prime
purpose of this firm which is the delivery
of highest quality DNA analysis of the most
modern standard to all concerned in industry, university, clinic and organizations for
private research.
6
Milagros Lazo Silva 2 Dr. Andreas Wolf
Martina Dreier 7 Dr. Bernd-Peter Ernst
12 Dr. Michael Betzler
13 Jasna Fraatz
1
3
8
Secondary structures, inverted repeats,
homo- and heteropolymeric stretches,
high GC or ATs …
We are not saying they are no problem but
we do say that, as sure as the “Gänseliesel”
of Göttingen is the most kissed girl in Germany, we can solve them. If you give us a
true sample of DNA for our Full Service we
will not rest until the sequence lies open.
Promise!
Dr. Werner Baussmerth 4 Dr. Mustafa Benli 5 Dr. Otto Knobloch
Katharina Dörnte 9 Dr. Rüdiger Kind 10 Dr. Karla Busch 11 Dr. Poulad Monzavi
04 SEQLAB
WHAT WE DO
We are DNA service providers and
have set as kernel of our competence the sequencing on commission in our differing, individually
relevant proffered services:
The HotShot is the ace in SEQLABs history.
Although fluorescent colour band sequencing has been available since the 90s and
was offered by various sequencing centres
the costs remained so high that many
smaller laboratories and university departments preferred to use their own resources. This involved radioactive methods as
a rule, and the yield, following two days of
heavy work, was information on a modest
250 bases. The breakthrough occurred in
1999 with SEQLABs HotShot – primer and
the DNA for sequencing were sent ready
mixed – and the savings in work time were
passed on to the client in the form of reduced costs. Thus sequencing within one‘s
owns four walls became even less attractive economically. Since it was not possible
to repeat the cycle sequencing a suitable
cycle sequencing (CS) protocol had to be
developed to enable the sequencing of
DNAs including those with simple secondary structures to be carried out successfully at the first try. This protocol has
been available since 2000 and constantly
delivers good results. 97 % of clean DNAs
with no ex-cessive secondary structures
deliver 99 % exact readings.
AdvantageRead is HotShot‘s big brother
and fills the gap towards the FullService.
Based on the same CS protocol it renders
none the less possible a repeat evaluation
under optimal conditions through a separate delivery of primer and DNA substrate
should the first attempt be inconclusive
and deliver poor results, due to unsuitable
secondary structures.
The FullService is a more advanced step.
Our experience at SEQLAB in chemistry
and the CS evaluation enables us to identify just about every sequence, thus bringing us to the reputation of ‘problem solvers’. There are virtually no DNAs with
secondary structures which find their way
to SEQLAB and remain undeciphered.
Apart from the sequencing described we
also offer a wide spectrum of further services and products:
GENOMICS
Nutcracker sequencing of hairpin DNA
structures; direct sequencing of cosmids,
fosmids, BACs and PACs, whole genome
sequencing, shotgun sequencing of larger
constructs; primer design and synthesis, in
vitro amplification, plasmid preparations;
DNA purification systems for many purposes;
PROTEOMICS
Protein sequencing and identification;
Polyclonal antibodies from various animal
sources; peptides with and without modifications.
www.seqlab.de
GENOMICS 05
OUR SEQUENCING SERVICES
Customer‘s work
SEQLAB‘s work
ca. 300 b
HotShot
9C 6
g
7.95 €
Eg^b
Z
ca. 900 b
+ BIGDYE
11.75 €
%#'ba"IjWZ
ca. 650 b
13.80 €
9C
6
AdvantageRead
24-h-Service
+ PRIMER
+ BIGDYE
ca. 900 b
16.80 €
Posted usually within 24 h
(+repetition if necessary)
25.50 €
9C
6
FullService*
S ca. 350 b
A ca. 650 b
+ PRIMER
+ BIGDYE
30.70 €
B ca. 900 b
36.90 €
* BACS, PACS AND GENOMIC PROJECTS ARE HANDLED AS FULLSERVICE.
Posted usually within 24 h
incl. basic editing (+repetition if necessary)
06 GENOMICS
YOUR ORDER (INFO AND TIPS)
The delivery of the sequence data follows usually 24 hours after receipt of
the post (24 hour service). The samples usually arrive before 12.30 pm,
are put into the cycle sequence reaction the same day and into the
sequencer that evening. Results are read and posted the following day.
HotShot sequencing
For the HotShot sequencing you send us
the template and the primer of choice (already mixed) – in a PCR tube of 200 µl with
a flat lid. The PCR cups should be numbered on the lid beginning with 1 and then
sequentially. Total volume should not exceed 7 µl: DNA in 10 mM TRIS, pH 8.5;
20 pmol primer. HotShot samples should
not be wrapped in parafilm or foil.
Necessary DNA
quantity
FOR HOTSHOT SEQUENCING
For plasmid DNAs please send 0.6 – 0.7 µg.
For PCR products the following quantities
should be sent: length of fragment divided
by 4 = quantity in ng, e. g.: PCR product
~500 bp : 4 = ~125 ng; a little extra is no
disadvantage. The primer should be one of
18 – 25mer with a Tm of 52 – 60 °C.
Please observe these exact instructions for
the HotShot sequencing. They are indispensable for the successful automatic work
programme.
FOR ADVANTAGEREAD SEQUENCING
Plasmid DNA: 1.0 – 3.0 µg with a concentration of 100 – 300 ng/ µl.
PCR products: quantity dependent on the
length with a concentration of: PCR fragment length divided by 20 = concentration
ng/µl; e. g.: PCR-Product ~500 bp : 20 =
~25 ng/µl.
FOR FULLSERVICE SEQUENCING
ds-DNA
(bp)
optimal
amount of DNA
(ng)
required concentration of
DNA (ng/µl*)
10000
2500
500,0
5000
1250
250,0
1000
250
50,0
750
138
37,5
500
125
25,0
250
63
12,5
FOR PCR PRODUCTS
As a rule of thumb: for DNA quantity: PCRfragment length divided by 4 = DNA quantity in ng; for the concentration: PCR length
divided by 20 = concentration in ng/µl.
For the sequencing we need 3 to 4 times
the amount of the calculated DNA quantity,
so that the possibility of repeating the sequence reactions is given, should need
arise.
FOR PLASMIDS
3 – 8 µg DNA depending on the size of the
vector. Minimum concentration is 200 ng/µl;
for larger plasmids (vector + insertion >
8000 bp) is a concentration of 300 – 400
ng/µl recommended. Please dissolve the
DNA in Tris/Cl, 10mM, pH 8.5!
GENOMICS 07
84.5 % GC - content
82.3 % GC - content
Precipitation of DNA
Please precipitate the DNA solely at room
temperature; the reagents employed
should also be stored at room temperature.
Salts tend to precipitate at lower temperatures, thereby disturbing the sequencing.
The DNA should in addition be well washed
– with at least the same volume of 70 %
ethyl alcohol, with which the precipitation
is undertaken. Please leave the DNA precipitate for 5 – 10 minutes fully under 70 %
ethanol; preferable is a second washing.
Plasmid preparations
For a specimen of plasmid DNA to deliver
excellent sequence results the following
points should be observed:
1) culture medium residues carried over in
the preparation lead to bad results. The
medium must be completely removed following the centrifuging down of the cells.
We recommend therefore the washing of
the cells in Tris/Cl-puffer solution (10mM,
pH 8.5) and then the complete removal of
the puffer.
2) Overloaded Mini-, Midi- and Maxi-Kit columns deliver comparatively large amounts
of DNA, the sequencing of which is, however, difficult or impossible. We recommend therefore the purification by column
of minipreps max. 2 ml, of midipreps max.
20 ml and of maxipreps at the most 100 ml
cell suspension. As an alternative for low
copy plasmids: up to 10 ml cell suspension
for a miniprep, washing of the cells with
Tris/Cl buffer (10 ml, pH 8.5), and then a
second miniprep with 10 – 15 µg plasmid
DNA from the first preparation.
PCR products
The following tests improve the probability
of obtaining good sequence readings from
PCR products:
a) It should be possible to reamplify a good
yield of the PCR products; should the PCR
product band only be weakly visible by the
reamplification then the sequencing will
itself be poor.
b) A product should result only with both
PCR primers; to test this you can lay down
two PCR reactions and use the forward primer in one and the reverse primer in the
second. Should a product result by one of
these tests, then the corresponding primer
was used as the forward and also as the
reverse primer (so called ‘one-primer
PCR’). We note this problem frequently
with PCR products of genomic DNA.
c) PCR products should be cleared of superfluous primers and dNTPs as well as of
any primer dimer products to avoid influencing the following sequence reactions.
OVER 90 AVAILABLE PRIMERS are listed on
our internet page under ‘services’, link:
‘available primers’. You can speedily find
there which primers are appropriate for
your vector. The use of these primers is included in our offer on your commission of
Advantage-Read or Full Service.
www.seqlab.de
Primerdesign/
-synthesis
Primers can also be ordered. Simple and
advantageous primers (up to 25mer) are
offered in conjunction to a sequencing
commission.
PACKAGING OF THE SAMPLES FOR TRANSPORT BY POST:
Please use padded envelopes for your
post. Since these are not a 100 % protection against breakage the tubes should be
additionally packed in plastic boxes, falcon
casings or small cardboard boxes. HotShot
samples should be solely sent in 200 µl
cups with a flat lid. These must not be
wrapped in parafilm or similar foil! The
DNA samples for Advantage-Read and FullService sequencing commissions should
be sent in 1.5 ml cups (the lid alone
wrapped with parafilm) or in 1.5 ml screw
cap vessels.
08 GENOMICS
YOUR DATA
1. Online service
SAFE, FAST, TRUSTWORTHY
Commission and delivery of sequence data
is usually by means of our server system
SEQLABdirect. To use SEQLABdirect you
only need your customer’s number and a
password. We inform you of your customer’s number following your first written
order. The password is sent automatically
via e-mail at the next commission.
In case of large amounts of data we will
compress the sequence files, in order to fit
more information into an e-mail. The compressed files have the ending *.sea. On
Mac-Computers they are decompressed
automatically by a simple mouse click. On
PCs you will have to decompress the files
with stuffit.
2. Other delivery
possibilities
Programmes for data editing
The following simple programmes for
observance and editing are on the
internet:
• for PCs (freeware):
CHROMAS Lite 2.01 from Technelysium:
http://www.technelysium.com.au/
chromas_lite.html
• for MACS (freeware):
EDIT VIEW from Applied Biosystems:
http://www.appliedbiosystems.com/
support/software/
For decompressing files:
• for PCs (freeware):
STUFFIT from SmithMirco Software
http://www.stuffit.com
The data can be sent per e-mail should you
so request. The four colour prints of the
electropherograms on paper or the delivery
of the data on a CD-ROM or a diskette is
possible for a slight extra charge (see current pricelist).
SEQLABS ONLINE SEQUENCES
You can also call up your sequence
results from any internet computer
without recourse to special software
(Chromas, EditView etc.). An exclusive
SEQLAB service.
YOUR ADVANTAGE
Prepaid HotShots
On subscription of Prepaid HotShots we
give up to 36 % discount.
Please ring us on 0551 - 370 00 - 11 to arrange your prepaid account. Following the
establishment of your prepaid account you
can only commission Prepaid HotShots online. Written orders are unfortunately not
possible. All other HotShot instructions are
unaltered.
0551 - 370 00 - 11
Dr. Werner Baussmerth
GENOMICS 09
OUR PURIFYING SYSTEMS
In the course of development of
the DNA preparation methods the
original phenol/chloroform DNA
extraction process according to
Maniatis was soon superceded by
procedures which function without phenol and which are based
Miniprep-Plasmid-Kit
on silicon- and other types of
The Miniprep-Plasmid-Kit
• delivers more quickly than a
conventional Plasmid-Kit
• a solid amount of
• highly pure DNA.
membrane allowing a selective
binding of DNA. These procedures
are justly regarded as having the
highest standard on the present
day market and as being mature.
Notwithstanding this we have
succeeded in raising the technical
standard yet again: a strict procedure entailing a much shorter
time of preparation delivers a
constantly high DNA yield of superb quality.
Clean DNA: excellent sequencing quality with single
and double repeat-motives
SLH-Low-CopyPlasmid-Kit
This Kit is used in the extraction of plasmid
DNA from low copy strains.
10 preparations,
article no.: 81010
18.– €
50 preparations,
article no.: 81050
75.– €
250 preparations,
article no.: 81250
280.– €
The SLH-Low-Copy-Plasmid-Kit
• delivers in low copy plasmids a solid
amount of
• highly purified DNA and
• combines the high yield of classic plasmid preparation methods with the
• high purity and extraordinary quality of
a process based on the affinity method.
10 preparations,
article no.: 82010
25.– €
10 GENOMICS
PCR-PURIFYING-KITS
PCR-Purifying-Kit
(2-STEP-PROCESS)
You use our PCR-Purifying-Kit for a fast
purification and concentration of PCR reactions in only two stages.
Adding the binding
buffer to PCR solution
1. Step:
Binding of PCR
fragments
2. Step:
Elution of
PCR fragments
Employ the advantages of the PCR-Purifying-Kit to enjoy the simple and fast
working method as well as the excellent
quality of the purified DNA.
With the PCR-Purifying-Kit it takes less
than five minutes to obtain a quick and
easy purification. The 2-step-process (binding and elution) is
• exceptionally suitable for fragments of
between 80bp and 30kb.
• The kit has flexible elution volumes,
down to a minimum of 10 µl at its disposal and is
• also suitable for the concentration of
samples.
The Gel-Extraction-Kit is used for a rapid
extraction of DNA fragments out of TAEand TBE-gels. In approximately one quarter
of an hour it achieves
• close to 100 % extraction and the
• highest quality and purification of
the fragment
The Gel-Extraction-Kit is the method of
choice where the pureness and unicarity
of the DNA is at stake.
10 purifications;
article no.: 83010
15.– €
10 extractions,
article no.: 84010
16.– €
50 purifications,
article no.: 83050
65.– €
50 extractions,
article no.: 84050
68.– €
250 purifications,
article no.: 83250
235.– €
250 extractions,
article no.: 84250
240.– €
Gel-Extraction-Kit
Those PCR products which appear in the
gel as a band may also be accompanied by
primerdimers (and other secondary products) in just non-visible concentrations.
These can interfere, due to thermodynamic
promotion, either with the cloning of the
PCR products, or cause double sequencing
of the product in the first 40 – 60 bases,
due to cosequencing of the dimer.
PCR-Combi-Kit
(PCR PRODUCT PURIFICATION OUT OF
SOLUTIONS OR GELS)
The PCR-Combi-Kit makes a highly efficient
purification of DNAs out of reaction solution and agarose gels possible. In both
operations the PCR-Combi-Kit delivers
• highly pure DNA of
• exceptional quality
10 purifications,
article no.: 85010
17.– €
50 purifications,
article no.: 85050
70.– €
250 purifications,
article no.: 85250
245.– €
GENOMICS 11
Plant-DNA-Kit
RNA-Mini-Kit
Bacteria-DNA-Kit
Our Plant-DNA-Kit enables up to 100 mg
of genomic DNA to be isolated from varying vegetable material. Following the
lysis of the vegetable substance with consecutive purification from unsoluble components and unwelcome contaminants
(e. g. phenols)
• binding of the genomic DNA by means
of a filter cartridge and the
• elution of highly pure DNA is achieved.
Our RNA-Mini-Kit enables the speedy and
efficient isolation of the total RNA out of
eukaryotic single celled organisms, bacteria and macerated tissue. Following the
lysis of the original substance a DNase I
free selective purification of the genomic
DNA is carried out. The RNA is then repeatedly washed and eluted. The obtained
total RNA is of
• excellent quality and
• outstandingly suitable for all further
applications.
Our Bacteria-DNA-Kit enables the speedy
and efficient isolation of bacterial genomic
DNA out of pellets of grampositive and negative bacteria from young cultures. Following the initial lysis step and the consequent proteinase K digestion of the proteigenic cell residues the liberated DNA is
bound to the membrane of the affinity filter cartridge. The bound DNA is then eluted
• with a good yield and
• in outstanding quality.
10 preparations,
article no.: 86010
50 preparations,
article nor.: 86050
250 preparations,
article no.: 86250
28.– €
108.– €
10 preparations,
article no.: 87010
30.– €
475.– €
50 preparations,
article no.: 87050
145.– €
250 preparations,
article no.: 87250
650.– €
10 preparations,
article no.: 88010
28.– €
50 preparations,
article no.: 88050
120.– €
250 preparations,
article no.: 88250
499.– €
0551 - 370 00 - 13
Dr. Otto Knobloch
according to ABI user protocol
Do you find this a hard nut to crack?
by SEQLAB
... just leave it to us!
12 GENOMICS
OUR FAVOURITE THERMOCYCLER
At SEQLAB we have worked with many
thermocyclers. From the first time on that
we tested the SENSOQUEST LabCycler we
were positive: this one and no other for us.
Its precision, quality and flexibility speak
for themselves since it combines in one all
those advantages that one has always
looked for in a thermocycler. In addition it
also looks good. Arrange for us to set one
up for trial in your rooms and see if it
doesn’t convince you: this also is your
thermocycler!
THERMOCYCLER
• Temperature: -5 °C to + 99.9 °C
• Heating rate: 4.2 °C per second
• Cooling rate: 3.2 °C per second
• Optional with gradient: 20 °C (+/- 10 °C)
• Autoresume function
• Heating lid: automatic, temperature and
pressure are programmable
• Electrical potential: 85 V to 265 V
without switching over
• Length: 444 mm, width: 251 mm,
height: 201 mm
LabCycler Basic
(without gradient): 011-103
5 875.– €
LabCycler Standard
(with gradient): 011-101
6 575.– €
0551 - 370 00 - 19
Dr. Bernd-Peter Ernst
USERS SURFACE
Languages: German, English
Protocol function
Intuitive service software
TFT touchscreen
Coloured graphic display
RS232 interface for software updates
•
•
•
•
•
•
Gradient upgrade:
011-801
700.– €
THERMOBLOCKS
• Electro formed gilded silver
• Processual steered gradient capable
block with individually steered peltier
elements for a particularly homogenic
temperature with high heating and cooling rates.
• Unique universal alternation block
system
• 48-wells block for individual tubes (0.5
ml)
• 96-wells block for microtitreplates 96 or
individual tubes (0.2 ml)
• 384-wells block for 384‘ microtitreplates
Thermoblock 48: 012-102
2 190.– €
2 190.– €
2 190.– €
PROTEOMICS 13
OUR POLYCLONAL ANTIBODIES
For the antibody production we work with
rabbits, mice, rats and guinea pigs. If you
wish you may also choose chicken, goat or
hamster. Place your order with SEQLAB informally, a special order form is not necessary.
The following sample protocol can be varied to suit your wishes: before the ‘immunisation’ has begun you will receive 2 to 5
pre immune sera which you can test for
possible cross reactions. You receive 2 ml
testaliquot after each blood withdrawal.
Standard immunisation scheme (3 months)
Standard immunisation scheme (2 months)
Day
0
1. Injection (preimmune serum >5 ml)
21
2. Injection
Day
0
1. Injection (preimmune serum)
21
2. Injection
35
1. Blood withdrawal (10 – 20 ml)
35
1. Blood withdrawal (10 – 20 ml)
49
3. Injection
49
3. Injection
63
2. Blood withdrawal (2 ml a. 10 – 20 ml)
53
2. Blood withdrawal (2 ml a. 10 – 20 ml)
77
4. Injection
60
Exsanguination
98
Exsanguination
ALSO ON OFFER:
• Antibody labeling (HRPO,
biotin, fluorescin)
• Quality check under enzymatic
THE NECESSARY ANTIGEN FOR ANTIBODY PRODUCTION IN RABBITS:
•
Form of antigen
1. Injection
Follow-up injection
Protein > 20 kD
Protein < 20 kD
100 – 500 µg
250 – 500 µg
100 µg
100 – 250 µg
Insoluble protein, microbodies
polysaccharides
50 – 250 µg
500 – 1000 µg
50 – 100 µg
500 – 1000 µg
Peptides linked to carrier molecules (BSA, OVA, KLH)
500 – 1000 µg
250 – 500 µg
Non viable bacteria, viruses, yeast protein
•
•
Please deliver antigens in physiological puffer solution (e. g. PBS, pH 7 – 8), lyophylised,
in a PAA sample or blotted onto nitrocellulose membranes.
Volume of antigen required: 500 – 1000 µl per injection.
•
control and HPLC/SDS-PAGE or
ELISA analysis
Antibody purification: affinitypurification by means of protein A- or protein G-sepharose
Affinity purification of sera by
means of an antigen matrix
Antibody fragmentation, FAB
fragments
ELISA test in a 96-wells
antigen microtitreplate
0551 - 370 00 - 11
14 PROTEOMICS
OUR PROTEIN IDENTIFICATION AND SEQUENCING
SEQLAB offers a mass-spectrometric (MALDI-TOF) analysis for the
measurement of the purity and
molar mass of proteins, as well as
for protein sequencing by the degradation method of Edman.
Protein identification
In the mass spectrometric analysis for the
measurement of purity and the molar
mass of proteins you can choose between
a total mass measurement and a fingerprint following trypsine digestion. Simply
send us a cut out SDS band in a cup. To
prevent desiccation add 2 drops of water
with a pipette.
Protein sequencing
after Edman
For the protein sequencing according to
Edman the individual amino acids are
split off and determined one after the
other.
OUR SERVICE COMPRISES:
• Chromatogram and process data of the
separate runs
• Log data for the individual amino acids
• Declaration of the determined amino
acid sequences
• Nomenclature and charge numbers of
the reagents employed, should this be
desired
REQUIREMENTS FOR THE SAMPLES TO BE
SEQUENCED:
• The samples should be lyophylised
• If possible, please send the relevant
chromatogram
• A definite coomassie band should be
visible in blot samples
• Sample quantity ca. 1 µg, or, for more
extensive commissions, up to 5 µg.
• Use solely PVDV membranes for blotting
• The sample should have as little as
possible contact with acetic acid, since
otherwise sequencing may not be
feasible.
HPLC-separation of PTH-amino acids
Delivery time: around 2 – 3 weeks
0551 - 370 00 - 10
Dr. Rüdiger Kind
PROTEOMICS 15
OUR PEPTIDE SYNTHESIS
Peptide syntheses are also offered
by SEQLAB. Synthesis is carried out
by the solid-phase-method under
the employment of the most
modern synthesis machines. We
proceed according to the FMOC
method.
Peptide synthesis
You may choose between different purity
levels:
Above 70 % peptide purity is sufficient for
the production of polyclonal antibodies
(‘immunisation’) by various animals. Purity stages above 80 %, over 90 % and over
95 % are necessary for more demanding
immune studies in vitro and in vivo. All
peptides are purified with HPLC. A mass
spectrogram is delivered with each peptide, thus enabling an exact identification
to be made. In addition we offer specific
modifications of the peptides such as
MAPS, MAP-4, D-aminoacids, N-acetylisation and C-amidisation.
Phosphorylisation with phosphoserine and
phosphothreonine can also be carried out.
Further modifications are available on request.
Conjugations on various carrier proteins
can also be performed. As a rule the conjugation of ca. 2 – 3 mg peptide and BSA or
KLH is undertaken. Please enquire as to
further carriers.
We check if the sequence that you request
involves changes in the standard synthesis
method on receipt of your commission.
Should these cause an increase in the cost
you will, of course, be informed immediately.
Delivery time
Peptide syntheses with HPLC purification
up to 20 AS are carried out within 4 weeks.
Longer peptides and such with modifications or ‘difficult’ amino acids can take up
to 6 weeks.
0551 - 370 00 - 11
16
GENOMICS PRICE LIST VALID FROM 20TH FEB. 2007, PLUS VAT
Order
no
40000
50000
40050
40100
40500
41000
50050
50100
50500
51000
40384
40096
50384
50096
60000
70000
60050
60100
60500
61000
70050
70100
70500
71000
10000
20000
30000
Service offer
Description and comments
Single tube HotShot sequencing
HotShot (read length ca. 300 b)
Extended HotShot (read length ca. 900 b)
Prepaid HotShot sequencing
50 HotShot < 300 bases
50 Extended HotShot < 900 bases
HotShot sequencing in microtitreplates
384 well HotShot ca. 300 bases
ADVANTAGE READ
Advantage 650 (ca. 650 bases)
Advantage 900 (ca. 900 bases)
PREPAID ADVANTAGE 650/900
50 Advantage 650 (length ca. 650 bases)
FULLSERVICE SEQUENCING
Short run S (ca. 350 bases)
Standard run A (ca. 650 bases)
Long run B (ca. 900 bases)
We can only guarantee these very low prices, if you exactly fulfil all the conditions: put the
appropriate amount of DNA dissolved in 5 – 10 mM TRIS- Cl, pH 8.0 – 8.5 plus 20 pmoles of the
primer in a total volume of 7 µl together in a 200 µl PCR tube with a flat lid. Label the tubes
only on the lid with consecutive numbers and don‘t wrap them with parafilm etc., but put them
in a stable box or plastic tube for protection and place this assembly in a shock-absorbing
padded envelope. We need 0.6 µg of plasmidary DNA and 25 ng for each 100 bases of length
of PCR-products (e.g. 50 ng of a 200 bases PCR-product). The resulting sequences are not
edited, the reactions can‘t be repeated. An invoice will be issued irrespective of the sequencing
result. Resulting sequencing data and electropherograms at SEQLAB Server. First order on the
HotShot order form only, then please use our ‘online service’ at www.seqlab.de.
Hot Shots in microtitreplates (MTPs) should be prepared same as above; customary MTPs schuld
be filled with components and covered with an adhesive film. Label the plate on the margin of
the MTP and on the film with a summarized name. The resulting sequences are not edited, the
reactions can‘t be repeated.
Cycle sequencing, universal or specific primers, capillary electrophoresis on the 3730 ABI
automatic sequencer; resulting sequencing data and electropherograms at SEQLAB Server.
The AdvantageRead closes a gap between the HotShot and our FullService Sequencing, the
latter of which offers 3 different read lengths (No. 1000 S to 3000 B). Although it includes an
extended treatment compared to the HotShot (i. e. repeated cycle sequencing with different
primer and/or DNA amounts), it falls behind the sophisticated trouble shooting of our
FullService Sequencing procedure. First order on the Advantage order form only, then please
use our ‘online service’ at www.seqlab.de.
Cycle sequencing with universal primers, gel elelectrophoresis, basic verification, sequence data
and electropherogram via e-mail. The resulting sequence is preliminarily edited. All Ns in the
start of peaks are removed from the sequence and the whole sequence is cleaned up from
gross irregularities by individual editing. The sequence data will be delivered to you by e-mail
with a short comment. All sequencing data are kept in the database for 3 months at SEQLAB,
allowing you the opportunity to order further walking beyond those delivered sequences.
Primer walking; cycle sequencing, electrophoresis, sequence data and electropherogram via
e-mail. The resulting sequences are fully edited, additional alignment at 99,99 % fidelity. The
expected sequencing error rate is 0.1 % per base for single strand primer walking and < 0.04 %
per base for double strand primer walking.
Cycle sequencing with universal primers, gel electrophoresis, basic verification, sequence data
and electropherogram via e-mail. SEQLAB has the rich know-how for sequencing large
constructs and can perform sophisticated sequencing protocols, leading to highly successful
sequencing of most BACs, PACs and genomes. Unfortunately there is no warranty for success
despite this optimisation.
By genomic direct sequencing is not possible to tell wich length were available.
Price €
7.95
11.75
Ask for prices
6.20/well
6.90/well
9.10/well
9.90/well
13.80
16.80
Ask for prices
25.50*
30.70*
35.90*
* Incl. OD- and
GC-extra charge.
70011
70012
70013
PRIMER WALKING
Single stranded sequencing
Sequencing of both strands in publication
quality > 1000 bp
BACS, PACS direct sequencing
BAC-S (ca. 350 bases)
BAC-A (ca. 650 bases)
BAC-B (up to 900 bases
70014
Direct sequencing of genomic DNA
70015
Genomic walking (in situ positioning)
70021
Primer design
70022
Primer synthesis
80000
Projects, genomic analyses
Shotgun sequencing, large inserts
80030
Plasmid preparation in MTPs
96 miniprep in microtitreplates
80031
Plasmid preparation
Miniprep
10.00
80032
Column purification
Using membranes for purification of plasmids, PCR products, Cosmids, BACs or PACs
10.00
80033
Purification of PCR products (Ag.)
Agarose gel purification or spin column
14.00
80035
Purification of PCR products (PAA)
High resolution gel separation and extraction. It is possible to discriminate DNA fragments with
only 2 bp length difference on polyacrylamide (PAA) gels.
25.50
70001
70002
0.10 p.b.
0.25 p.bp.
25.50
30.70
35.90
55.00
For the determination of the insertion locus of a known DNA section in its genomic surrounding, Ask for prices
or also for sequencing from a known DNA section into the unknown
At primer selection we receive its highest biological specificity by using modern primer design
18.00
programs with subsequent individiual critical examination of the proposed sequences. Our long
years of experience help us choose the right ones.
Primer, 10 nmol scale, up to 25mer primers
from 8.00 on
Price lists also available as PDF at www.seqlab.de
Ask for prices
2.00/well
GENOMICS PRICE LIST VALID FROM 20TH FEB. 2007, PLUS VAT
Order
no
Service offer
80036
Purification of PCR products in MTPs
96 well purification of PCR products in microtitreplates
80037
In vitro DNA-amplification
In vitro DNA-amplification
80041
80042
Quality control of DNA
OD (concentration and purity);
agarose gel or polyacrylamide gel (per fragment)
80047
DNA precipitation
With ethanol
3.50
80048
Cup-to-cup transfer
Transformation from unsuitable cups to suitable ones
2.50
80049
Concentrating of DNA
Concentrating of samples with < 200 ng/µl
2.50
80054
Sequence verification (full editing)
80055
Alignment
80073
Nutcracker
Firstly, this feature consists of the trimming of the raw data for the first uninterpretable
10.00/file
oversteered signals at the beginning of the sequence and those uselessly low signals at the end
of the sequence. Both kinds of signals disturb the computational integration of the desired
peaks. Then, we remove all of the Ns at peak start, which are derived.
From system immanent electrophoresis failures. Subsequently comes the careful examination of
10.00/file
the whole sequence including the manual correction of base calling failures and identifying of
real (biological) irregularities.
Attempt with special arrangement
from 250.– on
81010
81050
81250
82010
Plasmid preparation kit
83010
83050
83250
84010
SLH low copy plasmid kit
1.00/well
15.00
(*) is incl. in Full Service
18.00
75.00
280.00
25.00
15.00
65.00
235.00
16.00
80081
Delivery and packaging
80082
Delivery and packaging
Mail (abroad, without Antibody delivery)
80091
Set-up charge
80092
Set-up charge direct genomic sequencing
These charges will be invoiced only if the repeated sequencing attempts have failed even
under specialised conditions because your DNA was dirty or degraded or the primer was
wrong.
Only if the direct genomic sequencing has failed totally.
Gelextraction kit
PCR combi kit
Plant DNA kit
RNA mini kit
Bacteria DNA kit
HotShot cups
GC-special
Data transfer
80061
80062
80063
80064
3.50*
3.50
Kit for 10 minipreps via a new membrane for faster preparation of highly pure plasmids (P)
Kit for 50 minipreps
Kit for 250 minipreps
Kit for 10 low copy Plasmid preparations via a new membrane (P)
Kit for 10 PCR purifications with a new affinity based membrane in two steps (P)
Kit for 50 PCR purifications
Kit for 10 extractions via a new affinity based membrane in tree steps for faster preparation
of highly pure DNA
Kit for 10 DNA purifications from solution or gel in two or tree steps
Kit for 50 DNA purifications
Kit for 250 DNA purifications
Kit for 10 DNA preparations from plant cell for faster preparation of highly pure DNA
Kit for 50 preps
Kit for 250 preps
Kit for 10 RNA preparations with a new affinity based membrane
Kit for 50 RNA preparations
kit for 250 RNA preparations
Kit for 10 bacteria DNA präparations with a new affinity based membrane
Kit for 50 bacteria purifications
Kit for 250 bacteria purifications
500 PCR cups with flat lid for HotShots
1000 cups
GC-special for inverted repeats, GC extra rich sequences, sequences with particular tricky
secondary structures.
Attempt with special arrangement.
Mail (domestic, without Antibody delivery)
84050
84250
85010
85050
85250
86010
86050
86250
87010
87050
87250
88010
88050
88250
80102
80103
80072
PCR purification kit
Price €
Data delivery normaly at the SEQLABdirect server
Via diskette
Via CD-ROM
Via e-mail
Via four colour print
68.00
240.00
17.00
70.00
245.00
28.00
108.00
475.00
30.00
145.00
650.00
28.00
120.00
499.00
30.00
50.00
20.00
incl. in
Full Service
5.00/10.00
9.00
14.00
30.00
free
3.00
5.00
2.50
1.50/page
18
PROTEOMICS PRICE LIST VALID FROM 20TH FEB. 2007, PLUS VAT
Order
no
Service offer
Price €
90210
90211
Immunization of rabbit, guinea-pig, Polyclonal antibodies, raised in rabbit, guinea-pig, hamster;
chicken, hamster
2 months’ protocol with your peptide/protein as antigen.
as noted above in 3 months protocol
90212
Immunization of mouse
Polyclonal antibodies, raised in mice; 2 months’ protocol
116.00/animal
90213
Immunization of rat
Polyclonal antibodies, raised in rat; 2 months’ protocol
142.00/animal
90214
90215
Immunization of goat
Immunization incl. peptide
synthesis and coupling
Polyclonal antibodies, raised in goat; 2 months’ protocol. each additional month 125,00 EUR
Immunization of 2 animals with artificial peptides as antigen including peptid synthesis and coupling;
3 months’ protocol
650.00/animal
898.00
90216
Preimmune serum
Per animal and week
10.00/animal
90083
Deliveries for immunization
For all deliveries of an immunization project
35.00
90311
Peptide synthesis 70/10
10 mg, over 70 % pure, all peptides will be purified using HPLC.
18.50/aa
90312
20 mg, over 70 % pure, HPLC
29.80/aa
90313
28.00/aa
198.00/animal
258.00/animal
90314
45.90/aa
90315
39.90/aa
90316
58.80/aa
90317
48.80/aa
90318
69.70/aa
90321
90322
90323
90324
90325
90326
90327
Modification of peptides (B)
Modification of peptides (N10)
Modification of peptides (N20)
Modification of peptides (C10)
Modification of peptides (C20)
Phosphorylation of the peptide
with phosphotyrosine, phosphoserine or phosphotheronine
Biotinylation of peptides
N-acetylization
10 mg
20 mg
C-amidation
10 mg
20 mg
Phosphorylation
10 mg
20 mg
90328
Other modifications of peptides
Labeled peptide
90331
Conjugation
Coupling of the peptide with BSA or KLH, conjugation will be made with 2 – 3 mg peptide
155.00
90411
Protein sequencing up to 4 AA
N-terminal, Edman´s degradation per amino acid
208.50
90412
90414
Protein sequencing > 4 AA
Protein identification (MT)
N-terminal
Proteins are enzymatically cleaved; cleavage products are then MALDI-TOF analysed. Only proteins the
charactarizations of which are already deposited in databanks can be identified.
48.00/aa
150.00
90415
Setup cost protein sequencing
In case of missing results of protein sequencing
125.00
90416
Setup cost MALDI-TOF
In case of missing results of MALDI-TOF
30.00
20.00
33.00
30.00
46.00
130.00
260.00
Ask for prices
80.00
e-mail
Country
Department
Order number
or
date
diskette
CD-ROM
signature
e-mail for MAC *.sea
e-mail for PC *.zip
DATA TRANSFER
Preferably online delivery of data from our server! If desired otherwise, please tick the
following forms of delivery: (compressed if necessary)
Sequence Laboratories Göttingen GmbH · Hannah-Vogt-Straße 1 · D-37085 Göttingen · Postfach 3343, 37023 Göttingen · Phone: +49 (0) 551 - 370 00 10 · Fax: +49 (0) 551 - 370 00 12 · e-mail: [email protected] · www.seqlab.de
primers should be 18 to 20mers with a Tm from 52 – 60 °C.
Concerning different templates use the following amounts of DNA:
plasmids:
0.6 – 0.7 µg
PCR-product ~1000 bp:
250 ng
125 ng
50 ng
Please mix the necessary amount (s.b.) of DNA in 5 – 10 mM
TRIS/Cl, pH 8.5, and 20 pmol of primer in a total volume of 7 µl.
remarks
SEQLAB offers the highest quality (with a perfect DNA up to 99.9 % reading reliability)
and guarantees absolute privacy of the data.
number tubes on the lid successively with two-place figures (01 – 96)
Extended HotShot (app. 900 b) *
*) no mixed orders please: use separate order forms
Standard HotShot (app. 300 b) *
AMOUNT OF DESIRED HOTSHOTS
For HotShots please send us:
A 200 µl PCR tube with a flat lid, containing the DNA and the primer for sequencing.
(already mixed)
2
1
data
ATTENTION: We can only guarantee these very low prices if you exactly fulfil all the conditions mentioned above. In case of a formal mistake, you can appoint the samples as AdvantageReads, but
we can`t send them back. If the amount or quality of DNA or the primer are suboptimal, this may lead to a total failure of the sequencing reaction. However, payment for HotShot sequencing is
obligate irrespective of the result. If the category is not ticked, the order will be regarded as Extended HotShot order.
Don’t wrap any parafilm etc. around the tube, but put it into a stable box or plastic tube as protector and place this assembly in a wadded envelope.
Fill in this order form carefully and pay attention to primer and template concentrations and amounts!
Dear customer, please use this order form only for the first order, after that please use our online service at www.seqlab.de.
Fax
City
Street
Phone
Company/Institute
WWW.SEQLAB.DE
Name
ORDER FORM HOTSHOTS
Department
WWW.SEQLAB.DE
Company/Institute
ORDER FORM ADVANTAGEREAD
Name
Country
e-mail
City
Fax
Street
Phone
Order number
Dear Customer, please fill in this order carefully and pay attention to primer and template concentrations, use the same name on the tubes as in the order form! After purification please dissolve the DNA in 5 – 10 mM TRIS, pH8.5 (no EDTA)!
Wrap the tube tops with parafilm and send the samples well protected per mail. You need not send universal primers, but send specific primers in an extra tube.
length (kb)
reading length remarks
ATTENTION: The Advantage 650/900 holds a second attempt with optimised DNA/Primer-relation out. Please, order the analysis of more difficult templates on an extra order form as FullService.
Those templates will be treated intensively with specific SEQLAB-protocols (s. price list, No. 10000-30000 and 80072). NEW are the different lengths. Please mind that an unticked order will be processed as Advantage 900.
vector
900 b
template data
650 b
900 b
primer name
PCR
650 b
900 b
template name
Pl
PCR
650 b
900 b
please tick
1
Pl
PCR
650 b
900 b
vector/insert
PCR fragment
2
Pl
PCR
650 b
900 b
distributor
cloning site
3
Pl
PCR
650 b
please write in block letters, max. 6, but no
greek letters
4
Pl
PCR
kind, concentration, quantity
we need per reaction:
Pl asmid (100 – 300 ng/µl), 10 µl
PCR-Produkt (~bp-Länge/20 = ng/µl), 10 µl
5
Pl
Tm= 52 – 60 °C;
18 – 25mer;
10 pmol/µl, each 10 µl
6
More than 100 universal primers are available, e. g.: T3, T7 promotor, T7 terminator, M13 forward -21, M13 rev, SP6, U19, pGEX 3‘, pGEX 5‘, pGL 1, pGL 2
signature
DATA TRANSFER
Preferably online delivery of data from our server! If desired otherwise, please tick the following forms of delivery: (compressed if necessary)
diskette
CD-ROM
e-mail for PC *.zip
colour print
fax
date
e-mail
Country
Department
Order number
Pl
Pl
Pl
Pl
3
4
5
6
Cos
Cos
Cos
Cos
Cos
PCR
PCR
PCR
PCR
PCR
PCR
vector/insert
PCR fragment
length (kb)
sequence comparison
sequence alignment
colour print
date
fax
data base retrieval
signature
S
S
S
S
S
S
A
A
A
A
A
A
please tick
S ca. 350 b
A ca. 650 b
B ca. 900 b
B
B
B
B
B
B
high GC-content (> 60 %),
kind of preparation,
secondary structure
reading length remarks
DATA TRANSFER (compressed if necessary)
diskette
CD-ROM
e-mail for PC *.zip
sequence verification
More than 100 universal primers are available, e. g.: T3, T7 promotor, T7 terminator, M13 forw. -21, M13 rev, SP6, U19, pGEX 3‘, pGEX 5‘, pGL 1, pGL 2
Pl
2
Cos
distributor
cloning site
kind, quantity and concentration
we need per reaction:
Pl asmid (3 µg/200 ng/µl)
Cos mid (9 µg/600 ng/µl)
PCR-Product ~1000 bp (1 µg/50 ng/µl)
PCR-Product ~500 bp (500 ng/25 ng/µl)
PCR-Product ~200 bp (300 ng/20 ng/µl)
Pl
Tm= 52 – 60 °C;
18 – 25mer;
10 pmol/µl, each 10 µl
please write in block letters, max. 6,
but no greek letters
vector
template data
1
primer name
template name
ATTENTION: Difficult templates will be treated intensively with specific protocols developed by SEQLAB. This service is included since 01.01.2007
Dear Customer, please fill in this order carefully and pay attention to primer and template concentrations, use the same name on the tubes as in the order form! After purification please dissolve the DNA in 5 – 10 mM TRIS, pH8.5 (no EDTA)!
Wrap the tube tops with parafilm and send the samples well protected per mail. You need not send universal primers, but send specific primers in an extra tube.
Fax
City
Street
Phone
Company/Institute
WWW.SEQLAB.DE
Name
ORDER FORM FULLSERVICE
Company/Institute
Department
ORDER FORM PROTEIN SEQUENCING AND -IDENTIFICATION
Name
Country
e-mail
other:
2D
HPLC
sample in buffer (pH) purified with
(salt free sample)
other:
Elektrophoresis/Blot
HPLC
purified with
City
Fax
amount measured with
(Lowry, Bradford, BCA etc.)
µg
µg/µl
µg total
amount measured with
µg Lyophilisate
µg/µl
sample in buffer
(pH)
Street
Phone
molecular weight
PROTEIN SEQUENCING (EDMAN-DEGRADATION)
sample name
storage at
°C
molecular weight
PROTEIN IDENTIFICATION (MALDI-TOF)
sample name
storage at
°C
µg total
One data form for only one sample. (Order forms also available at www.seqlab.de)
date
WWW.SEQLAB.DE
Order number
assumed sequence
Data Base Research
Protein digestion
Mass Determination 1000 – 6000 Da
desired service
number of amino acids
blot/
staining technique
signature
CONTACT
Hannah-Vogt-Straße 1
D-37085 Göttingen
Postfach 3343, 37023 Göttingen
Phone
Fax
+49 (0) 551 - 370 00 10
+49 (0) 551 - 370 00 12
e-mail [email protected]
www.seqlab.de
administration, business management
Dr. Werner Baussmerth certified chemist
Phone: +49 (0) 551 - 370 00 - 11
e-mail: [email protected]
administration, business management
Dr. Bernd-Peter Ernst certified chemist
Phone: +49 (0) 551 - 370 00 - 19
lab management, business management
Dr. Rüdiger Kind certified chemist
Phone: +49 (0) 551 - 370 00 - 10
DNA diagnostics, business management
Dr. Otto Knobloch certified biologist
Phone: +49 (0) 551 - 370 00 - 13
marketing, business management
Dr. Poulad Monzavi certified agricultural engineer
Phone: +49 (0) 551 - 370 00 - 10
www.seqlab.de
Sequence Laboratories
Göttingen GmbH
Hannah-Vogt-Straße 1
D-37085 Göttingen
Postfach 3343, 37023 Göttingen
Phone
Fax
+49 (0) 551 - 370 00 10
+49 (0) 551 - 370 00 12
e-mail [email protected]
www.seqlab.de

SEQLAB – The best thing that can happen to a DNA.

Transcrição

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