DNA Sequencing Sample Preparation

To submit samples, you can either premix the template and the primer, or send them separately. The following are the conditions for both premixing and non-premixing sample preparations. When possible, please use 8-strip PCR tubes for less than 96 samples and primers, and 96-well plates for equal to or greater than 96 samples:

PREMIX Template with Primer (Premixed Samples)

    1. Premixed total volume (in water): 15 ul
    2. Premixed template quantity (in water):
        • PCR products:
                         • 100-200bp 2-3 ng
                         • 200-500bp 6-8 ng
                         • 500-1000bp 10-15 ng
                         • 1000-2000bp 20-30 ng
                         • >2000bp 60-100 ng
        • Single-stranded: 100-150 ng
        • Double-stranded: (plasmids) 500 ng
    3. Premixed primer (in water): 8 pmole

Submit Template and Primers Separately (Non-Premixed Samples)

    1. Non-premixed template quantity (in water):
        • PCR products: concentration > 10ng/ul, please send > 10ul and let us          know the concentration and the length of the PCR product.
        • Plasmids: concentration > 50ng/ul (100ng/ul is preferred), please send >          10 ul each.

    2. Non-premixed Primer (in water): concentration 3 uM, please send > 10 ul each, and have enough for all the samples.

BAC DNA and Genomic DNA Sequencing

Elim has extensive experience and provides BAC DNA and Genomic DNA sequencing. Results will take 1 more business day to obtain. When submitting samples for BAC DNA and Genomic DNA sequencing, submit in non-premixed form and please refer below for template and primer requirements:

    BAC DNA:
        • Template: >400 ng per sequencing reaction
        • Primer: 12 pmole per sequencing reaction

    Genomic DNA:
        • Template : 3-6 ug per sequencing reaction
        • Primer: 15-30 pmole per sequencing reaction
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