Samples must be purified after first PCR and delivered exactly in required concentration. Too low a concentration could lead to a low PCR yield, high concentration to different yields or even inhibition of the reaction (high DNA concentration may be associated with a high concentration of inhibitors). Please provide samples in 0,2 mL strips or 96-well plates.
It is required that samples are provided with tails (overhangs) as described, purified from artifacts such as excess primers or adaptors, enzymes etc.
Total lenght of DNA fragments must be between 200 and 900 bp!
How to design primers
Append to 5’ end of your primers:
Forward: 5’ TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-[locus specific sequence]
Reverse: 5’ GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG-[locus specific sequence]
Despite to coronavirus situation the sequencing was performed really quickly! The quality of reads is good. The facility at our institute is getting open again, so we won't request something from your side in the near future, but definitely we would recommend SEQme as the sequencing provider.
