Assay summary for RNA-seq_Giraldez_Lab_0002AS

Assay Type:
RNA-seq
Lab:
Giraldez Lab, Yale
Description:
For RNA input, total RNA was isolated from 400 ul of the 800 ul of clarified extract before ART- seq Nuclease treatment. Poly-A selection was done according to manufacturer guide- lines (Dynabeads mRNA Purification Kit, Cat no.610.06), and RNA fragmented using the Artseq Ribosome Profiling Kit Mammalian protocol. RNA input fragments were cloned according to the Artseq Ribosome Profiling Kit, Mammalian. The final PCR was carried out with an initial 15 second denaturation at 98 °C, followed by 9-12 cycles of 15 seconds at 98 °C, 5 seconds at 55 °C and extension at 72 °C for 10 s. Reactions were separated on a non-denaturing 8% polyacrylamide TBE gel and DNA fragments of the correct size were extracted and sequenced.
Library Preparation:
poly(A)+
Added by User:
giraldezlab
Date Added:
May 6, 2016, 9:36 a.m.
Last Edited:
Oct. 11, 2018, 7:08 a.m.

Applied in Series

Applied on Biosamples