Stephan Spangenberg

5 Questions 9 Answers 0 Followers

Questions related from Stephan Spangenberg

Can qPCR be successfully performed with low starting quantities of RNA (less than 100 ng)?

I have total RNA extracted from samples which are rather precious. I am trying to perform cDNA synthesis then qPCR. The RNA appears to be intact and reasonably pure, but I only have between 20 and...

03 March 2019 7,609 4 View

Over what temperature range will cell lines immortalized with temperature sensitive SV40 T antigen grow?

Hello Research Gate, I am planning on using a quite specific cell line which has been immortalized with the temperature sensitive SV40 T antigen. Cells immortalized this way are widely reported to...

01 January 2018 8,680 4 View

Can CRISPR/Cas9 Knockout be estimated by Western Blot before clonal isolation?

I have a CRISPR/Cas9 Wild Type (pSpCas9(BB)-2A-GFP (PX458)) plasmid that I have prepared, targeted for a nuclear protein's gene. I have already confirmed its sequence. I would like to test if it...

10 October 2014 1,227 5 View

How can I detect lentiviral particles in a solution?

I am attempting to produce lentivirus vectors using the pLKO.1 system according to the following protocol: http://www.addgene.org/tools/protocols/plko/ In this protocol, HEK 293T cells are...

07 July 2014 6,249 7 View

Why is my restriction digest producing overly large transformants?

I cut a MSCV vector with NotI and SalI. The vector is 6kb long and the restriction sites are about 100 base pairs apart. Upon performing gel electrophoresis, the cut vector produced a clear,...

06 June 2014 790 6 View