Nisha Manav

4 Questions 7 Answers 0 Followers

Questions related from Nisha Manav

Why did I get high gene expression in cells transduced with sgRNA cloned pLentiCRISPRv2?

I transduced ovarian cancer cell lines (A2780 and OVCAR4 ) with pLentiCRISPRv2 having sgRNA. after puromycin selection at 0.75ug/ml concentration for 8 days I isolated RNA and converted to cDNA...

28 April 2021 7,623 4 View

Can someone help me with the protocol for lentiviral transduction of plentiCRISPRV2 to generate knockout clones?

I have cloned my sgRNA in pLentiCRISPRv2 plasmid. I have tried lentiviral transduction of this plasmid into my ovarian cancer cell lines but the protein expression does not seem to alter. I am...

24 April 2021 4,577 6 View

Why my lenticrisprv2 plasmid after ligation of oligos is of smaller size than the expected?

after miniprep, I sequenced the plasmid. After sequencing I found that a region of plasmid was missing and its size if of around 8kb while it should be of around 12kb. It has happened to me for...

22 March 2020 930 3 View

Why my colony PCR gave false positive result?

I am cloning my guide RNA in pLenticrisprv2 plasmid. I picked colonies and did primary culture to miniprep. after miniprep i did a pcr using vector specific (hU6 promoter) forward primer and my...

22 March 2020 2,554 0 View