SERVICES OFFERED
DNA Sequencing:
The main service offered by the DNA Sequencing Facility is Sequencing Reaction and Electrophoresis. All DNA samples are sequenced using BigDye Terminator chemistry with available stock primers or user-supplied primers. The available stock primers are T7, M13 forward (-40), M13 forward (-21), M13 reverse, SP6, and T3. The default primer for M13 forward is M13 forward (-40). The DNA sequencing reactions are electrophoresed on ABI’s 3730xl DNA Analyzer which can produce read lengths of 1000 bases for high quality DNA templates in 1-2 business days. All sequencing reaction plates and individual samples must have acceptable quality controls before the results are released. The facility has an automatic pro-active redo policy whereby most poor quality reactions are repeated once for no extra charge. The facility will not re‑sequence samples that do not produce a detectable sequence signal during the initial sequencing reaction. The redo policy helps compensate for random reaction failures and also allows the facility to adjust the sequencing reaction for individual sample failures. When the samples have been sequenced the investigator will be notified by email. The email will have attached a compressed electronic copy of the DNA sequence as a text file. Also, a color printout of the electropherogram (trace file) will be placed in the investigator’s folder at the DNA Sequencing Facility. Upon request, the DNA text file and electropherogram file will be placed on a password secure FTP server.
Additional Services:
The DNA Sequencing Facility also provides a DNA fragment analysis service. The fragment analysis service is used for microsatellite genotyping, SNP genotyping and mutation detection. The fragment analysis is performed using ABI’s 3100 and 3730 DNA Analyzers. Pricing is dependent on the project size; please contact the DNA Sequencing Facility for more information.
Table 2. Optimal
DNA Concentrations
| DNA
Type |
DNA
Size |
DNA
Concentration |
| Plasmid |
3-10
kb |
100
ng/ul |
| Plasmid |
>10
kb |
100 ng/ul |
| PCR
Product |
<
1 kb |
50 ng/ul |
| PCR
Product |
1-3
kb |
100 ng/ul |
| BAC
DNA |
|
250
ng/ul |