Sequencing Sample Preparation
Samples with Primer and Template Separated
Template Type DNA Concentration DNA Volume Primer Concentration Primer Volume
Plasmid
50-100 ng/µl
10 µl
3.2 µM
10 µl
PCR(100-500bp)
2-5 ng/µl
20 µl
3.2 µM
10 µl
PCR(500bp-1kb)
5-10 ng/µl
20 µl
3.2 µM
10 µl
PCR(>1kb)
10-20 ng/µl
20 µl
3.2 µM
10 µl
Cosmid
0.5-1 µg/µl
20 µl
3.2 µM
10 µl
Fosmid
0.5-1 µg/µl
20 µl
3.2 µM
10 µl
BAC
0.5-1 µg/µl
20 µl
3.2 µM
10 µl
Genomic DNA
2-3 µg/µl
20 µl
3.2 µM
20 µl
Samples with Primer and Template Mixed
Template Type Amount of DNA Amount of Primer Volumn (DNA+Primer)
Plasmid
500 ng
9 pmol
15 µl
PCR(100-500bp)
10 ng
9 pmol
15 µl
PCR(500bp-1kb)
15 ng
9 pmol
15 µl
PCR(>1kb)
50 ng
9 pmol
15 µl
Separated DNA templates and primers are preferred.
Mixed DNA templates and primers can be submitted together. If the sample is to be sequenced in two directions, two
tubes or plates containing the same DNA and different primers should be submitted.
96-well plates preferred for samples 96 and more.
