This could be the result of submitting too much sample for sequencing and overloading the BigDye™ terminator chemistry. Make sure the submitted sample is between 15-200 fmol in a 3 µl volume. This corresponds to the following mass amounts:
Size of PCR amplicon | ng of DNA (in 3 ul sample) |
---|---|
100 bp | 1 - 12 |
500 bp | 5 - 60 |
1000 bp | 10 - 120 |
3000 bp | 30 - 360 |
5000 bp | 50 - 600 |