Thomas Mühlenberg

3 Questions 112 Answers 0 Followers

Questions related from Thomas Mühlenberg

What's the second band in my cDNA after RT-PCR and RNAse H?

Hello, I am trying to generate ssDNA from an RNA template via RTPCR and consecutive digestion of the remaining/duplexed RNA by RNAse H. The first part is easy, but if I run my ssDNA on a gel I get...

29 November 2018 1,395 0 View

Why does my CRISPR/Cas9 induced point mutation by HDR with a ssODN template not work?

Hello, I am trying to introduce a point mutation (A-->T) in my mammalian cell line (say 293T) and have it stably express the mutated protein. My sgRNA works fine because I get nice indels at...

12 January 2016 1,074 2 View

How can I linearize large amounts of plasmid?

I am facing a few problems linearizing my vector for electroporation: I want to linearize large amounts, say 50µg, of the vector (~14k bp), and have to use a rather expensive restriction enzyme...

02 July 2015 5,215 19 View