We have three 3130XL Genetic Analyzers set up for Sanger sequencing and fragment analysis. Different levels of service are provided, depending on your needs.
Level 1: $4.00 per sample; <24 hour turnaround
This is a load-only service for clients who perform their own cycle sequencing reactions using Life Technologies BigDye Terminator V3.1 Cycle Sequencing kits.
Samples must be submitted in 0.2 ml PCR tubes. If possible, they should be submitted in completed batches of sixteen. All samples submitted must be accompanied by a completed Sample Submission Form.
The amount of template DNA recommended to perform your own cycle sequencing reactions will depend on the quality of your product and the chemistry used. Highly efficient chemistries (such as BigDye V3.1) require less template. In general, we recommend using the following DNA concentrations shown below for best results depending on your template.
|PCR product: |
|Double-stranded (Plasmid DNA)||250-500 ng|
|Cosmid, BAC||0.5-1.0 μg|
|Bacterial genomic DNA||2-3 μg|
The facility recommended cycle sequencing reaction is shown below.
|Big Dye Terminator V 3.1|
|5X Buffer||2.0 ul|
|Big Dye Mix||1.0 ul|
|Primer (1.87uM)||0.5 ul|
The facility recommended standardized cycle sequencing conditions are shown below.
|Cycle Sequencing Conditions on MJ Research Thermal Cycler|
|15 seconds @ 95°C||Step 1|
|10 seconds @ 50°C||Step 2|
|4 minutes @ 60°C||Step 3|
|Go to Step 1, 35X|
Samples for submission must be numbered consecutively starting with number 1 on the top of the lid of your first sample tube with indelible ink.
Sequencing sample results are sent to the client within 24 hours using the email provided on the submission form. Sample files will have the standard (.ab1) extension.
If a sample fails to produce any readable sequence due to a technical error on our part, it will be rerun automatically, using the original sample at no additional charge.