Quantitate using a gel (nanodrop isn’t reliable with PCR products)
For small templates use 50ng PCR product (please clean it up first as left over primers can mess up sequencing) and 4 picomoles of primer (4uL of a 1uM aliquot)
For larger templates, it is recommended to go high as 150ng (for approx. 2kb). Otherwise somewhere between the minimum and this will likely work. Use 4 picomoles of primer (4uL of a 1uM aliquot).
Plasmids
For small samples (2-6kb) 600ng of product and 4 picomoles of primer (4uL of a 1uM aliquot) is recommended.
For larger samples use more template/product (7-8kb: 700-800ng recommended, 9-10kb: 900-1000ng recommended) with 4 picomoles of primer (4uL of a 1uM aliquot).
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