Dec 04, 2024

Public workspaceRNA-seq protocol

DOI
https://dx.doi.org/10.17504/protocols.io.bp2l6d98rvqe/v1
  • Renhe Luo1,
  • Michael Beer2,
  • Danwei Huangfu1
  • 1Sloan Kettering Institute;
  • 2Johns Hopkins University
  • IGVF
Michael Beer
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DOI: https://dx.doi.org/10.17504/protocols.io.bp2l6d98rvqe/v1
Protocol Citation: Renhe Luo, Michael Beer, Danwei Huangfu 2024. RNA-seq protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l6d98rvqe/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 04, 2024
Last Modified: December 04, 2024
Protocol Integer ID: 113700
Keywords: seq protocol rna, seq protocol for esc, seq protocol, rna, esc, seq, de differentiation, protocol, differentiation
Funders Acknowledgements:
NHGRI
Grant ID: U01 HG012051
Abstract
RNA-seq protocol for ESC-DE differentiation timecourse
Troubleshooting
RNA-seq protocol
RiboGreen quantification and quality control by Agilent BioAnalyzer.
500ng of total RNA with RIN values of 6.5-10 underwent polyA selection and TruSeq library preparation according to instructions provided by Illumina (TruSeq Stranded mRNA LT Kit; RS-122-2102), with 8 cycles of PCR.
Samples were barcoded and run on a HiSeq 4000 or NovaSeq 6000 platform in a PE50 run, using the HiSeq 3000/4000 SBS Kit or NovaSeq 6000 SP or S2 Reagent Kit (100 Cycles) (Illumina).