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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