Reaction Protocol for RNase HII (M0288)
The following is a typical reaction protocol for nicking at the site of a single ribonucleotide within a dsDNA substrate or for nicking an Okazaki fragment 5´ to the ribonucleotide adjacent to the DNA
- Resuspend 100 pmol of double-stranded substrate into 50 μl of 1X ThermoPol Buffer.
- Add 1 μl of RNase II and mix thoroughly.
- Incubate at 37°C for 30 minutes.
Note: To achieve the best results for a given substrate, it is often helpful to test more than one concentration of ribonuclease.