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Questions related from Roberto Valli
Hi everyone, I need to make CRISPRi (CRISPR intereference) with a gRNA designed not in the promoter, but in the middle of the gene in an intronic sequence and I am designing the dCas9 plasmid. I...
03 February 2023 904 0 View
Dear all, I am planning to make IVT with a plasmid I already have. The plasmid is composed by: ..........T7 promoter - cDNA of my gene - P2A - eGFP - bGHPolyA ............ Looking in the datasheed...
25 January 2022 1,291 7 View
Dear all, I looking if there is the possibility of DEB test (and relative optimal diepoxybutane concentration) on lymphoblastoid cell lines. That's why, besides the classical DEB test on...
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Dear all, I have to transfect lymphoblastoid cell line and HL-60 cell line with vectors expressing shRNAs. I have read that CMV promoter could not work properly with ahematological-derived...
07 October 2019 6,781 2 View
Dear all, I should knockdown genes of interest in human primary skin fibroblasts. What shoul I proceed? 1) Is it better siRNA or shRNA? 2) lipofectamine (es: RNAiMAX) or nucelofection (es:...
24 April 2019 2,143 7 View
Hi everybody! The whidely used method to quantify the various ribosome fractions (40S, 60S, 80S and polysomes) are by sucrose gradient and fractioning. Unfortunately this techniques uses very high...
19 March 2019 5,173 2 View
I need HL-60 as neutrophilic models as the cell line is near to diploid and I need to edit by CRISPR/CAS9. I tried to differentiate HL-60 with 1uM ATRA for 5-6 days but I reached only 5-10% of...
06 June 2017 4,716 4 View
