Ian Cartwright

7 Questions 11 Answers 0 Followers

Questions related from Ian Cartwright

Near diploid human cancer cell line?

I am currently looking for a near diploid human cancer cell line, such as HCT116 and U87-MG. Preferably XY with modal chromosome # between 40-50, "normal" DNA repair pathways, and able to easily...

07 July 2015 4,671 2 View

Has anyone encountered issues with MCF10a not incorporating BrdU?

Hello All, I am trying to do a simple fluorescence plus Giemsa stain/acridine orange differential stain for chromosomes. I have had great success in the past using MEF/CHO/and various cancer cell...

12 December 2014 875 2 View

Has anyone had an issue with Gibson Assembly not connecting a plasmid and a single insert?

Hello All, I am currently struggling with getting a Gibson Assembly to work. I am trying to insert a 2.5 Kb insert into a roughly 12 Kb plasmid (modified pLEX-MCS). I have had the G.A work in the...

12 December 2014 4,257 7 View

Nonspecific PCR products and faint bands with Q5 polymerase - any thoughts?

Afternoon everyone, I am currently using Q5 polymerase from NEB to add restriction sites to an insert in a pBabepuro plasmid (~5400 Kb vector and 2.2 Kb insert). I have designed the primers to...

10 October 2014 4,525 4 View

Does anyone have the sequence for the pBabepuro-MYC-ER plasmid?

Hello, Addgene does not have the complete sequence published. Also, does anyone know if myc is truncated on both the N and C terminus, a primer designed to target the start of the myc CDS,...

09 September 2014 1,453 1 View

Does anyone have a protocol for introducing I-SceI enzyme into a cell?

I have a plasmid with an I-SceI restriction site which I want to only cut once, which I do not believe can happen with an I-SceI expression vector.

11 November 2013 6,486 0 View

Western Blot staining issue, poor B-actin band

I have been running several western blots over the past few days and have had the same issue where large proteins seem to stain nicely where as the smaller (~50 Kda) stain very poorly. I am using...

06 June 2013 9,754 4 View