welll... busy at the lab as usual... kind of starting to enjoy labwork... haha.. like quite fun... everything is like a challenge... and i really want to do it and get the right results... sometimes, the results don't come out the way i want and i get really upset... but i take note of all the things that i did wrong and make it better the second time.. like casting agarose gel...
haha... the first few times i cast gel... will always have many small bubbles on the surface... and when they take photos of it... it will like affect the clarity of the results.... well... the small bubbles are caused by boiling of the agarose.... everytime i see the small bubbles appearing, i panic and stop microwaving the agarose.... but hahahaha... i have experience now!!!!
if it starts boiling (small bubbles appearing) just let it boil somemore.... when you microwave it longer, it will boil more vigoriously with big bubbles... but now the difference is that the big bubbles appear very fast and burst very fast as well... so you end up with a clear agarose liquid with no bubbles.... then add the EtBr inside, swirl to mix, and pour into the prepared erm.. casting equipment... haha... cast alot of gels this week... due to doing alot of CHIP and RT-PCR... so have to run the samples and stuff... also scraped alot of flasks this week... scrape until my hand like going to break... but... got this feeling of satisfaction..... haha... so i guess all the hard work is worth it....
hmm... like never do much western blot this week... aiyoh... but that day i saw eric harvesting the cells... he said it was for FACs and western... so i guess i will have to prepare a new membrane next week... aiyoh... still must prepare RIPA buffer, sonicate the cells, do protein assay to measure concentration, use prism programme to calculate amount of proteins needed for loading... prepare samples for loading, heat shock samples, cast sds gel, load samples, run gel, prepare membrane and filter paper, transfer proteins using transblot, stain membrane, block membrane, add primary antibody, incubate, wash, add secondary antibody, incubate, wash, add substrate, incubate, go dark room do exposure, develop film, get results.... phew!!!!!!!!!! very very long process.... haven't even go into details like washing cell pellet, spinning down, vortexing, etc.... still got to do side duties like prepare buffers, fill pipette tips etc.... so busy busy busy!!!! but i'm enjoying it!!!! :) omg... i will seriously be damn bored during lab practicals when i go back to school...
and yay!!! got a date later with cake and sarah!!!! XD
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