Mutagenesis reactions (2bp substitution and 14bp deletion) were performed by
using 1U TagMaster Enzymes and 20ng of 8.1kb template
plasmids under the conditions of 95¡ãC 5min, 18cycles of
(95¡ãC 20s, 60¡ãC 10s, 68¡ãC 4min). Without any treatment, the
products were directly transformed into TagMaster
competent cells or DH5¦Á competent cells.
50~60 clones from each plate were picked, cultured, minipreped and sequenced to verify the desired mutations, and the mutagenesis rates were thus determined.
The results showed that TagMaster competent cells achieved
significant higher mutagenesis rate than DH5¦Á competent cells. |