3,675
edits
Annealing complementary primers: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
m
no edit summary
Barry Canton (talk | contribs) No edit summary |
Barry Canton (talk | contribs) mNo edit summary |
||
| Line 14: | Line 14: | ||
*A simpler approach is to add the above mix in a PCR tube to a beaker of boiling water and just allow the water to cool down naturally. Most primers pairs with length less than 100bp should be fully melted at 100<sup>o</sup>C and certainly any non-specific binding should be melted at that temperature. | *A simpler approach is to add the above mix in a PCR tube to a beaker of boiling water and just allow the water to cool down naturally. Most primers pairs with length less than 100bp should be fully melted at 100<sup>o</sup>C and certainly any non-specific binding should be melted at that temperature. | ||
*Unless you have ordered your primers with 5' phosphate added you will probably improve the efficiency of any subsequent cloning steps by | *Unless you have ordered your primers with 5' phosphate added you will probably improve the efficiency of any subsequent cloning steps by adding the 5' phosphate using a protocol such as [[PNK Treatment of DNA Ends]] | ||
