Yuming Wang

5 Questions 9 Answers 0 Followers

Questions related from Yuming Wang

How can I increase CRISPR/Cas9 targeting efficiency?

Hi All, I am new at CRISPR/Cas9 field, I tried my first CRISPR KO cell line but it failed. I transfected CRISPR/Cas9-gRNA into HEK293T cells via lipofectamine 3000, after 48 hours, I extracted...

03 August 2015 6,226 3 View

Is it possible to make site directed mutagenesis in plasmids larger than 15 kb?

Hi All, I want to generate a mutation in plasmid, but my plasmid is around 16 kb, is it possible to do it? I am planning to design two non-overlapping primers containing the mutation site to PCR...

24 July 2015 2,739 3 View

Can anyone suggest a differentiation protocol for SK-N-SH?

Hi All, I have SK-N-SH cells I would like to differentiate, what protocol do you normally use? I can see that people do it in different ways, I want to know which method gives you the most...

01 October 2014 610 2 View

How do you extract good quality of RNA from frozen brain tissues?

Hi all, I had a bad experience on extracting total RNA from frozen human brain tissues. The PMI for all tissues is from 3 to 22 hours. I used the Qiagen RNeasy Lipid kit and tissueruptor to...

23 August 2014 4,599 2 View

Does anyone have problems recovering neuroblastoma cells from ATCC?

Hi all, Recently I got a frozen vial of SH-SY5Y cells from ATCC, I've been recovering them in a small T25 flask for two weeks, but they don't expand at all and look quite stressed. After I...

15 August 2014 8,155 20 View