Prepare template:

Poor template quality is the most common cause of sequencing problems. See Prepare Templates for more information. We assume that you use DNA of known concentration in PCR and purified templates in your sequencing reactions.

• Check DNA concentration, e.g. with NanoDrop ND-1000
• Verify success of PCR with agarose gel electrophoresis
• Purify sequencing template (e.g. ExoI-SAP, EXO-SAPiT)

SeqLab needs 6.6 µl of purified PCR product including 8 pmol sequencing primer to perform sequencing reactions.

For each reaction pipette:

• 5.0 µl of template (PCR or plasmid)
• 1.6 µl of 5µM sequencing primer
• Mix properly
• Seal plate and spin samples down

NOTE: Fill plates by columns: A1, B1, C1..
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(from BigDye Terminator v3.1 Cycle Sequencing Kit protocol)