Elizabeth Radley

7 Questions 6 Answers 0 Followers

Questions related from Elizabeth Radley

How to grow mammalian cells in two separate media within the same dish?

Is it possible to grow a single dish of adherent mammalian cells (e.g. HEK293) where the cells form a continuous layer but there are different media compartments? We are looking at cell:cell...

04 August 2023 2,762 1 View

In what order should coverslips be coated with PLO/laminin/fibronectin for neuronal culture?

We are attempting to differentiate hiPSCs to cortical neurons and will be growing them on glass coverslips. We have to triple coat the glass with poly-L-ornithine (PLO), laminin and fibronectin....

14 February 2023 8,655 0 View

Can iPSCs be thawed into a different media than the one they were frozen in?

My lab is starting a new line of iPSC culture (KOLF2.1j). The cell suppliers provide a maintenance culture protocol using StemFlex medium with RevitaCell Supplement (both Gibco) on Synthemax-II-SC...

25 October 2022 2,717 3 View

What is an acceptable level of helper phage contamination in a library?

My lab has been having issues re-amplifying a M13 phage library, which was donated to us from another lab. It yields binders that we have since validated, so there's no issue with the library...

13 September 2022 4,672 1 View

What H2O2 concentration for RNase precipitation?

I am attempting to recreate a protocol from a colleague's notes but I'm missing some key info. Unfortunately the colleague has since left so I can't ask them directly. We are using rabbit...

25 March 2022 5,125 5 View

What is the suitable pH range for assays using Ubiquitin-AMC?

I'm using ubiquitin-AMC as the substrate for deubiquitination assays with several different deubiquitinases. Normally I'd do the assay in 50 mM HEPES (pH 7.5) and 150 mM NaCl, which I know works...

12 August 2021 9,688 2 View

Why did my PCR work one day and not the next?

Can anyone help with why my PCR worked one day and not the next? I am attempting to PCR amplify my empty vector backbone ahead of Gibson Assembly and am using one backbone with two sets of...

30 March 2021 6,134 2 View