Menno ter Huurne

5 Questions 4 Answers 0 Followers

Questions related from Menno ter Huurne

Which Restriction Site to include in Linker region?

Hi all, I want to tag an endogenous gene with a fluorescent protein (at the 3' end). I'd like to include a linker region and a restriction site in between the protein and the fluorescent tag. The...

06 June 2019 3,294 1 View

Can I get high quality RNA from PFA-fixed cells?

I want to fix my cells with PFA, store them at 4°C, then sort them and isolate RNA for sequencing. Is this possible?

10 October 2014 3,804 19 View

What is the effect of fixatives on RNA quality?

I want to Isolate RNA for sequencing. However, I first want to fix my cells. I was thinking of using 1% paraformaldehyde, is this possible? Does this affect the quality of the RNA? If so, which...

07 July 2014 1,981 1 View

Which housekeeping gene should be used in gene expression during the cell cycle for normalisation?

I want to determine the expression of several genes during different phases of the cell cycle (synchronised cells) using RT qPCR. For that, however, I need a housekeeping gene, preferably >1, that...

06 June 2014 2,308 4 View

Two cell populations in propidium Iodide stained cells (flow cytometry)?

For cell cycle analysis I fixed my cells using 75% ethanol (1mL cold PBS + 3mL 100% EtOH (-20'C), dropwise), and stained them with Propidium Iodide (PI). It is supposed to be a homogeneous cell...

03 March 2014 6,162 3 View