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Sequencing Services

 
 

Template/Primer Submission Guidelines

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Single Stranded Plasmid:

15-25ng/ul

Double Stranded Plasmid

100-150ng/ul

PCR fragments:

 

     100-200bp

1-2ng/ul

     200-500bp

1.5-5ng/ul

     500-1000bp

2.5-10ng/ul

     1000-2000bp

10-20ng/ul

     2000bp 

10-25ng/ul

BAC / Cosmid Templates:

300-500ng/ul

Sequencing Primers:

3uM

All templates and custom primers must be combined into one tube per sequencing reaction. Samples should be submitted in numerically labeled strip tubes with strip caps (we recommend strip tubes from Island Scientific, P/N IS-435), or 96-well plates. The online submission form can contain detailed template and primer names. A minimum of 5ul of template and 3ul of primer at 3uM are needed per sequencing reaction. This allows us to repeat the reaction if needed. The T7(s), T3, SP6, M13 (-21) Forward, and the M13 Reverse primers are provided by the SBRI sequencing facility at no additional cost. Please indicate in the comments section of the online request form if you need house primers added.

All DNA and primers should be resuspended in water or Tris buffer. Custom primers should be 18mer or larger with roughly a 50% GC content. Melting temperatures (Tm’s) should be around 50 degrees Celsius.

*Combined template and primer should be submitted in PCR strip tubes with strip caps. Tubes with individual or attached caps will not be accepted. We recommend strip tubes from Island Scientific, P/N IS-435.

 

 

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