Purified DNA

Purified Plasmids: The quality of the DNA sequencing reaction depends on the quality of the purified DNA. Most DNA purification kits should provide DNA at a quality sufficient for sequencing.  DNA should be provided at a concentration of 100ng/µL in a volume of 10µL for a single read. If using multiple reads, add 5µL volume per additional read. If your sample is lower than 100ng/µL, the sequence reaction will likely still be viable, but the lower the concentration, the lower quality the results may be. Please note on your order form the exact concentration if you have it, this will help us optimize the amount of template in the sequence reaction. Feel free to use this Dilution Calculator to help get the right concentration.

PCR Products: The quality of the DNA sequencing reaction depends on the cleanliness of the PCR product. In our lab we use an ExoSAP enzymatic clean-up, but other commercially available cleanup protocols will work as well. To avoid any degradation of the ends of the PCR products, we recommend cleaning the samples immediately after the PCR reaction. Purified PCR products should be provided in a total volume of at least 10µL and concentrated per the chart below. Feel free to use this Dilution Calculator to help get the right concentration. Any dyes left over from the PCR reaction are okay and should not interfere with sequencing quality.

Length of PCR amplicon Desired ng/µL

Sample and Primer Premixed: For pre-mixed samples we require a total of 20 picomoles of primer per 15µL of premix. We recommend using the following formula to create pre-mix samples:

DNA: N=0.1 x (vector size in base pairs + insert sizes in base pairs)

*N is the nanograms of DNA that should be added to each 15ul of premix

Do not premix primers into crude PCR products that still need PCR cleanup.

Samples sent as purified DNA will be sequenced at the following price: