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Keck
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DNA Sequencing
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Premixed Reactions
Individual tube submission policies and
procedures
Individual tube submission
policies
Individual Tube Submission
Protocol
Standard turn around time
individual tube submission
policies
1.)
all samples must be submitted with
primer and template premixed, except for primer walk projects.
2.) We only accept individual tubes
submitted in VWR Labcon and American Bioanalytical purple screw cap tubes.
The American Bioanalytical tubes are located in both Yale stockrooms.
Individual tube premixed reaction submitted in any other tube types will not be
accepted. The
American Bioanalytical tubes can be
order by phone at 1-800-443-0600 or through email at
info@americanbio.com. The catalog number is
PL73310-00001 and they are
packaged 500 tubes with caps.
3.) Due to our sequencing methods all
individual tube samples should be labeled with barcodes prior to submission.
Barcodes are given out over the counter in the Kline and Medical Stockrooms at
no additional cost. Please see Barcode
Labeling Instructions for more information.
INDIVIDUAL
TUBE SUBMISSION PROTOCOL
 Provide primer and template
premixed according to the guidelines below. We offer a high volume service
for large projects of 48 or more samples submitted at a time. Please see
High Volume Submission Policies and Procedures for
more information.
The protocols below are sufficient
for two sequencing reactions (e.g. 300ng DS DNA/seq.rxn.) with one
template/primer combination. This allows us to repeat a sample in case of
technical failure without having to contact you for more sample.
DS
plasmid DNA:
PCR Product:
e.g. 500bp PCR fragment, mix 25-50
ng PCR product with 2 µl 4 µM primer and sufficient water to bring the final
volume to 18 µl.
Cosmid, BAC and Phage
Lambda DNA:
Specify Cosmid, BAC or Phage DNA
on sample sheet so we will use altered cycling condition.
Only high purity templates should be used to optimize the
amount of DNA loaded onto the sequencer.
BAC template preparation methods that we recommend: 1)
Alkaline lysis with phenol extraction and isopropanol precipitation. 2)
Cesium Chloride Banding
Single-stranded M13 or
Phagemid DNA:
STANDARD TURN
AROUND TIME
Our turn around time is 24 -48 hours from the time we
receive your samples (except for samples received on Friday). For example,
samples received in the morning by 10:30 am will be returned the next business
day before noon.
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