Primer
Extension Sequencing (24 hour turnaround time):
Single
pass sequencing (24-hour turnaround) accommodates project
sizes ranging from single tube reactions to 384-well plates
and employs the same state-of-the-art sequencing technology
used in our HIGH-THROUGHPUT
SEQUENCING SERVICE. Samples may be submitted
as cells (glycerol stocks, stabs, plates) or DNA (plasmids,
PCR products, BAC/PAC, lambda, cosmids/phosmids). Service
highlights include:
· Automated
fluorescent dye-terminator sequencing on ABI Prism™ 3730xl
sequencers
· Universal primers provided (custom synthesis available)
· >650bp read lengths usually achieved
· Phred analysis of data (results returned to customer)
· Data returned as electronic transmission, diskette or hardcopy printout |
Customers
may either provide sequencing primers or provide an electronic
copy of primer sequences to be synthesized using SeqWright's OLIGO
SYNTHESIS SERVICE. SeqWright also provides--free
of charge--unversal primers common to most commercial vectors.
Some of the more common universal primers offered by SeqWright
include: T7, T7 Terminator, T3, M13F, M13R, SP6, BGH Reverse,
pGEX 5' and pGEX 3'. (See the COMPLETE
LIST OF UNIVERSAL PRIMERS provided by
SeqWright.)
Samples
being submitted for sequencing should be sent according
to the the guidellines outined in the following table:
Template
Type
|
Template
Concentration |
Template
Volume |
Custom
Primer Concentration |
Custom
Primer Volume |
| PCR |
30
ng/µl |
10 µl/reaction |
3.2 µM |
10 µl/reaction |
| Plasmid |
0.2 µg/µl |
10 µl/reaction |
3.2 µM |
10 µl/reaction |
| Cosmid,
BAC |
1 µg/µl |
10 µl/reaction |
10 µM |
10 µl/reaction |
| Genomic
DNA |
1 µg/µl |
10 µl/reaction |
10 µM |
10 µl/reaction |
GC
Rich, AT Rich and Difficult Templates:
SeqWright's
methods allow for clear delineation of base calls through
difficult templates. When standard dye-chemistry techniques
are unsuccessful, a variety of approaches including
PCR, transposons, nested deletions or small insert
libraries (SILs) may be selected to complete the project.
Our methods have been proven successful with the following
template types:
· GC-rich
and 2° structures (AAV ITRs)
· AT-rich (mtDNA)
· Homopolymeric tracts (cDNA PolyA, PolyG )
· Repetitive regions (PolyTG)
· Heterozygote insertions & deletions (Hetero-Indels) |
Ready-to-Run
(Load & Run):
For
a minimal cost, customers may submit ready-to-run
reactions for analysis on ABI Prism 3100/3730xl DNA
sequencers. Sequencing reaction clean-up is offered
at an additional charge. Data is returned to the
customer within 24 hours. The following dye chemistries
may be submitted:
· BigDye
Terminator v.1, v.1.1, v.3, v.3.1
· BigDye Primer v.1, v.3
· ET Terminator |
Feature
Reporting :
Automated
sequencing provides the most specific means of mutation
detection available. SeqWright’s proprietary
sequencing chemistry enhances mutation detection
protocols allowing rapid and consistent analysis
of large numbers of samples. SeqWright can provide
a final report annotating the locations of all mutant
bases. This report is not offered automatically but
is available upon request. Please visit our Resequencing/SNP
Detection page for more information.
Pricing:
Pricing
and turnaround times differ with template types and project
sizes. Discount pricing is available for large volume
orders (96-well to 384-well). Please contact our Sales
Department for
more information on this and other services.
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