Guidelines for DNA Sample Submission for Quick Single Pass DNA Sequencing Service

We understand that every sequencing result and turnaround time is critically important for you to proceed to the next step of your research. In order to generate good sequencing data the first time, please review the following information for details on how to submit DNA samples to ACGT, Inc.

Amount of Template and Primer DNA Required: The amount of template and primer required in each reaction is shown below. Please send around 10µl DNA to do a re-run in case the first reaction fails.  In a separate tube for template and primer: Template DNA Size of DNA Amount (ng in<5µl) Primer (pmol) Plasmid DNA up to 10 kb 200 10 PCR DNA 100-200 bp 10 10 PCR DNA 200-500 bp 20 10 PCR DNA 500 bp-1 kb 40 10 PCR DNA 1 kb-2 kb 100 10 PCR DNA over 2 kb 200 10 Large DNA over 10 kb 1000 20  Pre-mixed template & primer in the same tube: Please be sure to include at least 10µl of the pre-mixed samples. Template DNA Size of DNA Amount (ng) Primer (pmol) Total Vol(µl) Plasmid DNA up to 10 kb 100-200 20 10 PCR DNA 100-200 bp 10 20 10 PCR DNA 200-500 bp 20 20 10 PCR DNA 500 bp-1 kb 50 20 10 PCR DNA 1kb-2 kb 100 20 10 PCR DNA over 2 kb 200 20 10 Large DNA over 10 kb 500-1,000 40 20		General Information: 1. Samples can be submitted in microfuge tubes (e.g. 1.5 ml, 650µl, or other tubes) or 8-strip microtubes with the appropriate amount of template and primer. 2. Different samples submitted in 8-strip microtubes should have clear indications corresponding to template descriptions to avoid sample mix-up. 3. Extra amount of DNA ensures that we have enough samples to do a re-run in case the first reaction fails. 4. OPTIONAL: Please include a gel photograph of the template DNA with the quantity of the DNA loaded including a molecular marker. 5. Quantitation of template DNA - We recommend using gel electrophoresis, where the band intensity of a sample DNA is compared to a standard ladder, as the most accurate quantitation. For example, the 1.6 kb band of the popular Invitrogene 1 KB plus ladder contains 8% of total amounts of DNA present in the ladder mixture, and can be used as quick reference for estimating the concentration of your template. 6. Note that if a template requires to be analyzed in two directions, pre-mixed samples of the template with forward and reverse primers in separate tubes should be submitted. 7. Templates and primers should be clearly identified on the Sample Submission Form. 8. Please use a BLACK marker only to label your tubes. Colored markers smear and can be very difficult to read.