Jump to content

Elisa2804

New Members

  • Joined

  • Last visited

View Profile Find content

Everything posted by Elisa2804

  1. Elisa2804

    RNA Dephosphorylation

    Elisa2804 posted a topic in Biochemistry and Molecular Biology
    Good evening everyone, I'm working with sequencing of tRNA, applying the mim-tRNA Seq protocol (Behrens et al., 2021) but i don't understand the step of dephosphorylation, in which T4 PNK is employed... I know for sure that this step is crucial for adapter ligation on the 3' end of RNA, since when we have tried the ligation without a preliminar dephosphorylation step of total RNA and de-acylation step of tRNA, the ligation was failed. This enzyme has a 3'-phosphatase activity, but tRNA presents a 3'-OH end so, why is this dephosphorylation step important? I supposed that maybe during the de-acylation step (using Tis-HCl) a phosphoryl group remains attached on the 3'-end of tRNA, and so for this reason dephosphorylation is required, but i'm not sure. Can someone give me the answer to my doubt? Thank you in advance, Elisa

Important Information

We have placed cookies on your device to help make this website better. You can adjust your cookie settings, otherwise we'll assume you're okay to continue.

Configure browser push notifications
Chrome (Android)
  1. Tap the lock icon next to the address bar.
  2. Tap Permissions → Notifications.
  3. Adjust your preference.
Chrome (Desktop)
  1. Click the padlock icon in the address bar.
  2. Select Site settings.
  3. Find Notifications and adjust your preference.