Sarah Bell

6 Questions 7 Answers 0 Followers

Questions related from Sarah Bell

Lack of qPCR reproducibility- is it my cDNA?

Hello, I have been unable to replicate any of my qPCR samples no matter what I try. I am using the BIO-RAD qPCR machine with Sybr Green, and my own designed primers made using the ORFfinder...

11 November 2017 2,670 5 View

Is lipid transfection of mammalian cells with small quantities of plasmid DNA possible?

I am planning to transfect my mammalian cells with plasmid DNA but for various reasons, I only have a small amount (about 6 ul of 100ng/ul). I had planned to try a transfection on a 6-well plate...

05 May 2017 9,770 4 View

Validating a gene knockout?

I was wondering if anyone has any advice on validating a gene knockout? I have a cell line which has my gene of interest knocked out with a 1bp insertion into an exon. I have tried qPCR which was...

12 December 2016 8,244 4 View

Very high 260:280 when nanodropping RNA, what does this mean?

Hello, I'm using the RNeasy micro RNA micro kit from Qiagen to extract RNA from 3 samples. I've never had any problems with RNA extraction in the past, but for some reason I'm now having some very...

03 March 2016 6,855 4 View

Are you familiar with the puromycin concentration in HEK 293 Cells for shRNA?

Hello, I was just wondering if anyone had done shRNA with lentivirus in HEK 293 successfully and if so, what concentration of Puromycin was used? I've done a Kill Curve on a 6 well plate and I...

05 May 2015 8,393 8 View

How do I transfect siRNA in HEK 293 cells?

Hello, I was wondering if someone could help with siRNA transfection. I'm trying to lipid transfect HEK 293T cells using Qiagen's hiperfect transfection reagent. I have been working from the...

05 May 2015 2,328 7 View