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Questions related from Richard A Kahn
We are using CRISPR/Cas9 to introduce small indels to KO genes in immortalized MEFs and are finding that in almost all clones and different gene KOs that the null cells transfect very poorly. This...
19 June 2019 4,094 0 View
A graduate student recently asked me this question and I stumbling to give a good answer. We freeze our purified proteins quickly (liquid nitrogen or dry ice/EtOH) to prevent ice crystals from...
12 September 2018 5,624 3 View
The problem is that the activity is all in the P1 (pellet after homogenization and centrifugation at 1,000xg). When we apply the re-suspended P1 to a sucrose step gradient all the activity is at...
21 July 2017 8,955 0 View
I've tried five times now, using PCR to amplify and add 4 different combinations of restriction sites to try to insert into a pCDNA3 derivative. I've had great success with the vector with other...
13 July 2014 4,904 6 View
We grow human/mouse cell lines in culture (HeLa, HEK293, MEFs) in the absence of antibiotics. We regularly stain cells on coverslips with DAPI to test for mycoplasma. Lately we have seen what...
11 June 2014 8,521 6 View
