SOE PCR (Splicing by Overlap-Extension): Revision history

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10 August 2023

  • curprev 13:2113:21, 10 August 2023‎ EricMiller talk contribs‎ 755 bytes +755‎ Created page with "Eventually, a 50ul PCR. * 5x GC buffer (or whichever PCR buffer used): 10ul * Templates: Up to 100ng of largest fragment ** Equivalent molarity for all other fragments * dNTPs: 1ul * Water:To 47.5ul * Phusion: 0.5ul * PCR for 15 cycles "using the annealing temperature of the homologous regions". * Extension time: use the length of the (n-1) longest fragments, where n is the total number of fragments * Immediately after these 15 cycles: ** Add 1ul each of outside prime..."
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