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  • Reaction Protocol for RNase HII (M0288)

    Overview

    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

    Protocol

    1. Add 100 picomoles of double stranded substrate to 1X ThermoPol Buffer.
    2. Bring the reaction volume to 20 µl with water.
    3. Add 1 µl of RNaseHII and mix thoroughly.
    4. Incubate at 37°C for 15 minutes. 

      Note: To achieve the best results for a given substrate, it is often helpful to test more than one concentration of ribonuclease.