I had very few amount of my target Gene purified by gel extraction now to amplify gene I did pcr of the same....
so can I use this Pcr product directly use to my restriction digestion reaction..
or I need to gel do gel extraction again to elute out rest pcr master mix
usually you can cut directly in pcr buffer but New England Biolabs website has a useful page showing the percentage expected cutting for many enzymes in pcr buffers and the ability of enzymes to cut close to an end of the amplimer (for cloning ) will also depend on how many additional bases there are after the cut site. Again NEB have a list of additonal bases needed for efficient cutting
https://uk.neb.com/tools-and-resources/usage-guidelines/activity-of-restriction-enzymes-in-pcr-buffers
- Badges
- Science topic