Overnight Sanger Sequencing

Plasmid Samples

Quantity and Concentration Requirements:

1. A standard Sanger sequencing reaction requires approximately 200 ng of plasmid DNA.
2. The minimum volume for a plasmid DNA submission is 10 µL, with a minimum concentration of 30 ng/µL.
3. For plasmids exceeding 20 kb in size, the DNA quantity should be doubled to ensure optimal sequencing results.

PCR Samples, Purified and Unpurified

Sample Types:

We accept both purified and unpurified PCR samples. In the case of unpurified PCR samples, we will conduct a PCR purification process prior to sequencing.

Quantity Guidelines:

1. Approximately 10 ng of DNA per kb is required for PCR products.
2. Please provide the PCR product size information at the time of order submission.

E. coli Colonies

Acceptable Formats:

We accept E. coli samples in the form of agar plates, colony suspensions, or overnight cultures.

DNA Amplification Methods:

1. To prepare for sequencing, we employ one of the following DNA amplification methods:
   1. PCR (Preferred): This method allows for next-day data delivery. Please provide the amplicon size, as well as forward and reverse primers.
   2. Rolling Circle Amplification (RCA).
   3. Plasmid Miniprep: Include antibiotic information for the sample.
2. Note: RCA and plasmid miniprep methods may extend the data delivery timeline by an additional day.

Sample Packaging for Bacterial Suspensions:

Ensure all bacterial suspension samples are placed in either tightly sealed 8-strip tubes or in a strip-capped 96-well plate.

Genomic DNA

1. PCR amplification is necessary prior to sequencing. Please provide the PCR primers and the expected amplicon size.
2. A minimum of 10 µL of DNA sample is required, with a concentration of at least 30 ng/µL.