Collect amplified samples, new tubes (excluding negative control), and exoSAP enzyme solution kept on ice. Label all new tubes clearly for organization.
▶ 00:04
2
Transfer amplified samples to new tubes
Transfer 5 microliters of each amplified sample into labeled new tubes. Return the original amplified samples to the fridge.
▶ 01:27
3
Add exoSAP enzyme and incubate
Add 2 microliters of exoSAP enzyme solution to each sample, wiping the tip before expelling due to viscosity. Cap, mix, centrifuge to collect, then place in thermocycler on exoSAP program.
▶ 02:38
4
Prepare new tubes for sequencing reactions
Obtain two new tubes per digested DNA sample to create separate forward and reverse sequencing reactions. Label sample numbers on the front and F or R direction on the back.
▶ 06:10
5
Set up sequencing reaction master mix
Add 10 microliters of water to all new sequencing reaction tubes, then add 3 microliters of digested DNA sample to both its forward and reverse reaction tubes using fresh tips for each transfer.
▶ 08:16
6
Add primers and finalize reactions
Add 2 microliters of the appropriate primer (forward or reverse) to each corresponding reaction tube. Cap all tubes, mix, and centrifuge to collect samples.
▶ 09:30
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