Large Template

For templates larger than 10KB, please provide 1µg of template with 10pmol of primer in total volume of 13ul.

In addition, please write "Large  Template Protocol" in large print across the order form.

Cosmid, BAC, PAC, Fosmid

We are testing a new sequencing protocol for cosmid, BACs, PACs, and Fosmids.  We will sequence your sample for free of charge. If you have a  protocol you want us try, we will be happy to try it.

Please provide:

  • 5µg template with 30pmol of primer in total volume of 12ul.
  • Write the Tm of your sequencing primer on the order form.


Things to consider when preparing Cosmid, BACs, PACs and Fosmid for sequencing.

Template w/ secondary structure

If you suspect that there may be a secondary strucure in your template, please indicate in BIG RED MARKER that you want "dGTP PROTOCOL"

dGTP protocol may also work with GC and GT-rich templates.

Information

We accept samples in 310 Barker Hall all day from 9am to 8pm, Call 510-642-6383 after 5pm

We collect samples at 11:30 am 6:30 pm at the following locations:

Free Stock Primers

Free Stock Primers

M13Forward(-20)(17-mer): 5'GTAAAACGACGGCCAGT-3'

M13Reverse (24)(16-mer): 5'-AACAGCTATGACCATG-3'

T7Forward(20-mer): 5'-TAATACGACTCACTATAGGG-3'

T7Reverse(19-mer): 5'-GCTAGTTATTGCTCAGCGG-3'

T3(17-mer): 5'-ATTAACCCTCACTAAAG-3'

SP6(19-mer): 5'-GATTTAGGTGACACTATAG-3'

PCR Clean Up

PCR Clean Up:

By Plate:  $36/plate

NOTE: We elute PCR products in 40ul of H2O.   Enough for 6~7 sequencing reactions.

1.  For PCR reactions that consist of a single product and PCR products over 100bp.

2.  Up to 95 samples (we need well A1 for control).

3.  Please indicate on your order form the volumes of your PCR reactions.

Genotyping

Genotyping:

  1. Before you start your project, please make sure that you use the dye sets compatible with our Applied Biosystems 3730XL DNA Analyzers.
  2. The dye set best suited for our 3730XL is DS-33, which includes these dyes: 6-FAM, VIC, NED, PET,  and LIZ (LIZ Size Standard).  Please call us if you must use other dye sets.
  3. To submit your samples, you will need to use a 96 well half skirted or semi skirted PCR plate.  

    We prefer samples to be submitted in a specific type of plate:

Full Plate Submission

Full Plate Submission

You may also submit samples in a 96-well plate. We charge per sample, so you don't pay for empty wells. Please keep well A1 empty for our positive control.

Single Tube Submission

Please follow these guidelines when preparing your samples:

Each sample tube (1.5ml tubes preferred) should contain one primer and one template at the quantities specified below:

Please note that the values listed for the primer are quantities, not concentrations. 

  • ssDNA plasmid:  0.25μg DNA/5kb    + 2 pmol primer in 13 µl
  • dsDNA plasmid:  0.5μg DNA/5kb      + 8 pmol primer in 13 µl
  • PCR products:    2-4ng DNA/100bp  + 8 pmol primer in 13 µl

 

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