Sean Seltzer

4 Questions 33 Answers 0 Followers

Questions related from Sean Seltzer

For gene expression, is it better to average all the exons or is it better just to take the expression of a more stable region like the 3'UTR?

Some techniques involve averaging exon expression levels, but this can be a problem for genes that exhibit alternative splicing, so I'm wondering if there's a more stable region to take to get a...

23 June 2017 6,224 3 View

Is it biased to map RNA-Seq reads to a single chromosome instead of the whole genome?

I'm just interested in a single gene and don't need all the mapping to other chromosomes, and it may save me time. But is it biasing the reads to fall on sequences that are not the best matches...

20 January 2016 9,201 3 View

How do you do an RNA-IP while preserving RNA quality over the incubation period?

I use a RIPA lysis buffer containing RNAseOUT 1:2000 I cross-link the cells with 1% formaldehyde to stabilize RNP complexes, but this has a disastrous effect on RNA quality (though on the odd time...

04 May 2015 247 1 View

In transfected cells I can amplify GAPDH but I can barely detect the transgene, despite being detectable by western blot. Anyone understand why?

HeLa cells (6-well) transfected with 4ug pDNA Cells are lysed with NP40 Lysis Buffer with RNaseOUT Protein isolated from lysates shows expression of transgene by WB RNA isolated from lysates with...

08 April 2015 3,548 4 View