Dec 12, 2024
- 1Heinrich-Heine Universität Düsseldorf
Protocol Citation: Igem Dusseldorf 2024. cDNA synthesis. protocols.io https://dx.doi.org/10.17504/protocols.io.14egn9676l5d/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: December 11, 2024
Last Modified: December 12, 2024
Protocol Integer ID: 114727
Funders Acknowledgements:
Aligning Science Against Parkinson's
Grant ID: ASAP-020527
Abstract
Protocol for cDNA synthesis from RNA.
Materials
200 ng of RNA
preferred cDNA synthesis Kir
for cDNA synthesis, we use the BioRad iScript cDNA synthesis Kit (Cat.# 1708891)
use 50ng/uL of RNA for reaction, up to 1 ug total RNA per reaction volume (20 uL)
thermal cycler program: priming for 5 mins at 25C, reverse transcription for 20 mins at 46C, inactivation for 1 min at 95C, and an optional hold step at 4C.
used directly or stored at -20° C