13 Questions 9 Answers 0 Followers
Questions related from Lee Siggens
I have found in preliminary tests that many (6 tested so far) of the commercially available H4K20me3 antibodies can be blocked in ChIP assays by K20me2 peptides as well as K20me3. Has anybody...
27 March 2019 4,679 1 View
When performing ChIP-proteomics/ChIP-MS based techniques, if one chooses to do X-ChIP using standard 1% formaldehyde cross-linking is it necessary to reverse the cross-links before performing...
03 October 2018 1,112 3 View
What comes first in the DDR out of PARP activation and ATM/ATR mediated phosphorylation of H2AX? Is PARP required for the appearance of yH2AX or vice versa (i.e. ATM/ATR/yH2AX activate PARP)?
27 September 2017 4,625 2 View
Apart from fibroblast cell lines like IMR-90 etc, are there any good human cell lines that are not derived from a tumour. I'm interested in cell lines that can undergo quiescence cell cycle arrest...
11 March 2017 2,417 12 View
What do people commonly get as a transfection efficiency using plasmids for commonly used AML related suspension cell lines? Is 50% good? Does anyone have any special tips and tricks?
24 February 2017 4,683 2 View
Is it a guarantee to have a loss of expression or protein function if you have introduced an indel into a gene? In other words does a 1-3 bp indel definitely introduce a frameshift mutation or can...
24 February 2017 4,295 2 View
In Thurman et al (DOI:10.1038/nature11232) they report that 580 000 distal DHS sites were paired to TSSs. I cannot find this predicted DHS-promoter data-set from ENCODE. Presumably there is a big...
04 October 2016 7,392 0 View
In one recent study (Agirre et al 2015, Genome Research) the authors suggest B cell enhancers are inactivated in multiple myeloma. In stem cells/progenitors cells the aforementioned B cell...
07 April 2016 2,728 2 View
I was wondering if there are any genetic markers, especially deletions, in the U2-OS cell line that I can use to check if a sequencing data set is valid.
27 November 2015 1,629 2 View
We want to stimulate PNT2 or other prostate cells with DHT to stimulate AR dependent gene expression. What is the best way to do that and where can one buy cell culture grade DHT?
06 May 2015 1,604 1 View
Could someone tell me - when performing nascent RNA-seq in human (or mammalian) cells is it necessary to remove ribosomal RNA during library prep? Has anyone done this with Invitrogen's Click-IT...
10 September 2014 9,656 7 View
Usually there are slight differences in the protocols when we use DNase or MNase qPCR/ seq in analysing genome accessibility. Is it known if DNase sensitivity and MNase sensitivity (i.e....
12 May 2014 288 1 View
I have successfully knocked down several genes of interest using siRNA and LNA gapmers but have one particular gene that seems to be resistant (only one siRNA out of 7 and no LNA gamers worked)....
05 March 2014 7,968 3 View
