TO DO: *Culture up the ligated parts (J37022 & J37025) for miniprep (to check whether ligations from Monday were successful) from plates in incubator *Make frozen stock of 4G(S01656), J37018 and colony 2 of J37020 *Make more 50 times TAE *Culture for testing: T9002, J37015, J37020 (colony 2) *To order: LB medium (or make up ourselves & send to autoclave) & make some LB Amp to have in stock Autoclaved flasks (and cuvettes?) Need more eppendorfs More LB Amp plates ? (How many more do we need for our remaining time - there are about 10 left) *Gels from the weekend need to be analysed: Were J37018 & J37024 ligations successful ?
- Run once more, this time with DMSO, and a larger amount of maxiprepped DNA
- Major breakthrough - discovered that the reason for the repeated failures maybe that the DNA is there, but is not showing up.
- Today's gel was looked at immediately after being run and two distinct bands could be seen - however these disappeared once left for a few minutes.
- Once immersed in Ethidium bromide solution, the bands reappeared under the UV light.
- Repeat ligation J37023(Immunotagged AiiA)->1I
- Digest DNA from Maxi prep stock
- Run Maxi prep gel
- File:Maxi j37024.pdf
- Glass Milk Purify insert and vector (J37023 and 1I respectively)
- Leave to ligate overnight
Culture Up bacteria for testing tomorrow.