This is from memory 12hrs after the meeting so let me know if there are errors
- Dress code
- It was mentioned that Stephanie's glasses can draw attention from other things.
- Steph will make a spreadsheet with schedule for cleaning TC room.
- Cell line/Contamination status
- Leo's HEK293t/EGFP are OK.
- Steph libraries seem OK.
- Yvonne had trashed her old lines and started from scratch.
- Chase's HEK293t/EGFP were moved from MEM to DMEM/F12 and there was observed loss of fluorescence.
- These can be difficult to detect as they cannot be seen in the microscope but may be the cause of inconsistent experimental results. Their presence can also be masked if antibiotics are used, as signs of other contamination (e.g. Bacteria) would not be apparent yet mycoplasmas may still be present.
- Leo ordered primers for 16S rDNA PCR and will work on using this method.
- we had in the lab a kit including Hoescht DNA stain and primers to detect it. should be in one of the small freezers.
- Establishing line with a stably integrated protein
- Yvonne wanted to stably integrate an shRNA into some lines.
- Chase explained it would be best making a construct containing ZeoR and the region to be integrated containing as much of both as possible to avoid removing important regions e.g. Kozak sequence.
- Leo brough up using lentiviral vectors especially if cloning the same construct into multiple lines. However virus prep may be different for different lines, says Chase.
- Steph mentioned sometimes flip-in flips out :-).
- wiki site
- Leo brought up having a mammalian page on the wiki site which people can "watch" for updates or use to have a record of issues e.g. contamination.
- Next meeting: Monday August 21 at 2pm.