Hectic day

Well came in at usual time today - 9am. Then it was striaght off to image my gel... er i left it over night and all the DNA had diffused out!! Apparently i should have stored the gel wrapped in cling film in the fridge! Oh well i will know next time! Gel ran fine, there was cut plasmid. Digested my PCR inserts and used alkaline phosphatase to remove phosphates from the vector so it wont religate (the insert did not get dephosphorylated therefore can get ligated into the plasmid - remember AMB!). Also because all our supervisors leave at 5 (they all get in REALLY early around 8ish) we have to leave at 5 as well..... I was allowed to run my gel until 5:30, Steve said he would wait, but i left to talk to marv and forgot about my gel, I think i was a few mins late, therefore Steve wasnt happy with me, and i got a ticking off. He had to be somewhere... he ended up leaving... opps!

Karen, Xiao and Marv went to Gloucester road for lunch, would have loved to join them BUT had to be back in labs early (30mins time). It was a really quick lunch without having to wait for EVERYONE to be ready....

After lunch it was more running gels, and then exising the flourescent bands using a UV machine and scrapel and then extracting the DNA from the gel. I had a hige panick when i speced my DNA and found there was no reading! Nothing. What happened to my DNA. I was thinking just great, i dont make any mistakes and it all works, BUT during the important parts i muck up! I would have to repeat all this weeks worth of lab work!!!! I was not a happy bunny. I was advised to re-spec them, however before i did, Derek noticed that i was NOT using the UV cuvettes! Lol, THATS why there was no reading! When i came to specing them i had WAY too much DNA, so i had to dilute them down, otherwise i would be adding 0.03microlitres! Too small an amount! I am leaving the cut plasmid and insert to ligate at 4 degrees in the fridge overnight. Apparently the enzyme works best in room temp. BUT becasue of too much kinetic energy in room temp the insert and plasmid dont stay long tog. to get ligated. The best method is to use low temp overnight or 16 degrees for 2 hrs (compromise). Left at 5:30. And went stright home without going to comp room!