Overnight Sanger Sequencing

Plasmid Samples

1. About 200 ng plasmid DNA is used in one Sanger sequencing reaction, the maximal volume of plasmid DNA used is 5 ul. We prefer plasmid with minimal concentration of 30 ng/ul.
2. If the plasmid size is more than 20 kb, please double the amount of DNA.

PCR Samples, Purified or Unpurified

1. We accept both purified and unpurified PCR samples. If samples are unpurified PCR, we will perform PCR purification before sequencing.
2. For PCR product, we use about 10 ng DNA per kb, so please kindly provide PCR size information when making order.

E. coli colonies

1. We accept agar plate, colony suspension, overnight culture.
2. We will need to use one of these three methods to amplify DNA before sequencing. PCR is the first option we use, since this will allow us to deliver data the second day. RCA and plasmid miniprep will takes an extra day.
• PCR: please provide amplicon size, forward and reverse primer.
• Rolling Circle Amplification.
• Plasmid miniprep: please provide antibiotics information.
3. For bacterial suspension, place all samples in either tightly sealed 8-strip tubes or strip capped 96-well plate

Genomic DNA

1. We have to perform PCR amplification before sequencing, please provide PCR primers and expected amplicon size
2. Please send at least 10 ul DNA sample, with concentration ≥ 30 ng/ul