Table 2 Components of PCR solution for splicing of different numbers of DNA fragments
From: Simultaneous splicing of multiple DNA fragments in one PCR reaction
1Components | Two | Three | Three | Four | Five | Six | Seven |
|---|---|---|---|---|---|---|---|
Volume (μl) | |||||||
Total | 30 | 30 | 30 | 30 | 30 | 30 | 30 |
ddH2O | 15.7 | 14.7 | 14.7 | 13.7 | 12.7 | 11.7 | 10.7 |
2Buffer | 3 | 3 | 3 | 3 | 3 | 3 | 3 |
dNTPs | 3 | 3 | 3 | 3 | 3 | 3 | 3 |
Primer1 | 3(D1F) | 3(D5F) | 3(D2F) | 3(D4F) | 3(D3F) | 3(D2F) | 3(D1F) |
Primer2 | 3(D2R) | 3(D7R) | 3(D4R) | 3(D7R) | 3(D7R) | 3(D7R) | 3(D7R) |
Pfu | 0.3 | 0.3 | 0.3 | 0.3 | 0.3 | 0.3 | 0.3 |
D1 | 1 | – | – | – | – | – | 1 |
D2 | 1 | – | 1 | – | – | 1 | 1 |
D3 | – | – | 1 | – | 1 | 1 | 1 |
D4 | – | – | 1 | 1 | 1 | 1 | 1 |
D5 | – | 1 | – | 1 | 1 | 1 | 1 |
D6 | – | 1 | – | 1 | 1 | 1 | 1 |
D7 | – | 1 | – | 1 | 1 | 1 | 1 |