Sebastián Montealegre

3 Questions 4 Answers 0 Followers

Questions related from Sebastián Montealegre

Is there a good protocol to nucleofect Cas9+sgRNA RNPs into BJ1 hTERT cells?

I would like to nucleofect human BJ1 hTERT immortalised fibroblasts with a RNP complex made of Cas9 and a single crRNA+Tracer RNA, using the Amaxa 4D cell nucleofector device. Is there a standard...

02 March 2022 2,571 0 View

Problems doing a ligation into the lentiviral vector pLVX-EF1α-IRES-Puro?

I have been trying to clone mCherry from another vector into pLVX. mCherry is flanked by a NotI site at the 5' end and a BamHI site at the 3' end. The two sites are present into pLVX contigous one...

23 June 2016 8,015 4 View

Is it necessary to pre-treat HEK 293T cells with Neomycin before using them as packaging cells?

I want to use HEK 293T cells for the production of lentiviral particles. I was told that after thawing the cells from the liquid nitrogen, I should split them 4 times in the presence of 1 mg/ml...

12 August 2015 8,976 3 View