Irene Simkin
Core Manager
(617) 638-4096
(617) 638-5331
gencore@bu.edu
DNA Sequencing Requisition
A) Please fill in all information.
Date:
Your name:
Principle Investigator:
Telephone:
P.O.# or Evans Fund:
Email (to receive results):
Authorized Signature:
Billing Address (institution, room, street address, zip code):
Shipping Address (department, institution, room, street address, zip)
If BUMC
If BU Main Campus
If BMC
Other: (Please Specify)
B) Please circle the type of sequencing service desired and the size of the cloned
Plasmid
PCR
BAC/PAC
96 Plate
Run Only
insert or PCR product
C) Use 200ul PCR tubes and enter sample names below. Circle “IN” if you have already added your own primer or “ADD” and
specify one of our standard primers  M13 forward, M13 reverse, T7, T3, SP6, pGEX5’, pGEX3’. Total volume should always be
6ul for plasmid or PCR template, whether or not you add primer. Total volume for BAC or PAC template should be 24ul.
Plasmid insert PCR
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Plasmid insert PCR
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D) Bring this sheet with your samples attached to Evans 604 and put both in the Core freezer.
Project:
Date Completed:
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List Prices for Basic Core Services
I Florescent Sequencing
Sequence & run, per samples:
Plasmid and PCR preps
$10.00
II Florescent Genotyping
Run only, per gel, up to 96 lanes:
$175
 Volume discounts and other services available, just ask the Core Manager.
Sample Preparation
Submit your samples following the directions below. The quantity and quality of DNA greatly impacts sequencing success; too much
or too little leads to poor results. Tape your sample tubes to the requisition form, and put everything in the Core freezer at Evans 604.
Your sequence data will be emailed to you. Free software available through our website (http://gencore.bumc.bu.edu/), EditView for
Macs and Chromas for PCs, will display sequence chromatograms from the data files.
Sequence and Run  Use 200ul tubes
A) Plasmid template: Please supply purified, double stranded plasmid DNA in 6ul water. Do not use buffer. Provide 50ng DNA
for every 1Kb of template (vector and insert combined). Add 2 picomoles of your own primer or specify one of our standard
primers. The total volume should always be 6ul.
B) PCR product template: Please supply well-optimized, double stranded PCR product in 6ul water. Do not use buffer. Provide
50ng for every 1Kb of product. Add 2 picomoles of your own primer or specify one of our standard primers. The total volume
should always be 6ul.
C) BAC or PAC template: Please supply .5 to 1ug purified, linearized DNA (BAC only) in 24ul water. Do not use buffer. Add 20
to 40 pmol of primer. The total volume should always be 24ul. Large DNA templates are notoriously difficult to sequence.
Contact the Core Manager if you have questions.
Run only
Please suspend reactions with 5ul formamide loading buffer in a 1.5 ml microcentrifuge tube.
OR
For large sample numbers use 96-well format microtiter plates (or strip tubes) compatible with MJ Reasearch PCR blocks.
Policy Note
We use standard kits and widely accepted protocols with a control reaction run on every gel. If through the direct fault of the Core
sequencing fails, your template will be sequenced again at no additional charge. However, customers who request additional
sequencing free of charge should be prepared to demonstrate clean, robust template of appropriate concentration through a gel image
and spectrophotometer reads.