Template/Primer Submission Guidelines
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Services
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Single Stranded Plasmid: |
15-25ng/ul |
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Double Stranded Plasmid |
100-150ng/ul |
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PCR fragments: |
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100-200bp |
1-2ng/ul |
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200-500bp |
1.5-5ng/ul |
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500-1000bp |
2.5-10ng/ul |
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1000-2000bp |
10-20ng/ul |
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2000bp |
10-25ng/ul |
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BAC / Cosmid Templates: |
300-500ng/ul |
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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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