So the plan for the past few days was to keep my cells growing (enough to use them soon if they grow quick enough!), do a bit of monocyte work and set up and maintain biofilms.. and it has once again proved to be a somewhat challenging but rewarding week at that.
Cells have established themselves very well indeed, and I’ve managed to grow enough to have two sets of freezer stocks, and two flasks actively growing, and the keratinocytes are doing pretty well too 🙂 (albeit significantly slower growing than the fibs!). Monocyte work didn’t really go to plan..as ever..but still, learning how, and having the confidence to use the flow cytometer on my own is somewhat an achievement as far as I’m concerned – a mere microbiologist using some fancy tech haha! Still haven’t got the result I was hoping for, but it seems as though it is getting close to being statistically significant in it’s own right..so watch this space!
Back to the biofilm work that I’ve missed for some time while developing my ‘skills’ in tissue culture.. I fixed some for later (CLSM) and took some for RNA extraction, using the same method, but pooled three biofilms together..and managed to get more than 10x as much RNA yield!! Purity is amazing..but yield is just incredible. Considering for the past 2 years I’ve been getting yields of 40-80 ng/ul, to get 500 as the lowest is a pretty good achievement!! Fingers crossed for the next lot of batches too! I’ll have more DNA than I know what to do with 😀 (at least now I can do some proper qPCR optimising too..primer checks and all that jazz with consistent samples of DNA). Super chuffed to say the least!
Next week is the last before preparations for OMIG begin (9th-11th March, Gregynog (so excited)), and I have medic students with me Monday pm and Wed (am), Thursday and Friday all day..should be fun (I think!). Anyway, today is Thursday which means Friday tomorrow, and I’m looking forward to spending a lovely day with my two boys at the museum, Ruben is really looking forward to seeing the dinosaurs, and Oliver will be too I’m sure if he knew what was going on! Fun times ahead 😀