Saturday, August 2, 2008

2nd Week of Lab

Just finish my 2nd week of lab.
Starting to get a bit bored, especially when there is a lot of waiting time.
-waiting for the gel to harden
-waiting for gel electrophoresis to finish running
-waiting for PCR
-waiting for centrifuge machine to stop spinning
-waiting for seed to germinate and grow
-waiting for ........

still can't foresee myself doing this for the rest of my life..
Doing FYP is starting to get a bit 棘手.
Things no longer run as smooth as the year 1,2,3 experiments.
Designed and ordered my primers last week.
It arrived last Friday, so have to postpone the whole set of experiment to the following week.

Just wanna express my sienness here. So skip the next big chunk of more technical stuff, which will bore you.
Monday (28/7/08)
PCR, using my JmjC primers with Arabidopsis thaliana Columbia DNA as template.
Electrophoresis of the PCR products.
Gel extraction of the PCR products.

Tuesday (29/7/08)
Purify my PCR product.
Clone the PCR product into TOPO cloning vector.
Transformation of the competent cells.
Culture the cell, on Kanamycin selective plate.

Wednesday (30/7/08)
Pick the correct colony, and inoculate in LB broth.

Thursday (31/7/08)
Plasmid extraction from the bacteria cells.
Run restriction test.

Everything was surprisingly smooth so far...
THEN.... shit happened...
I used EcoRI to run restriction test, cos my gene was flank by EcoRI sites in the vector.
but the gel turn out to have a lot of small bands.
=( not the result I want.
Bo pian, went back to check the 5 genes, only to realize that some of the genes got a few internal EcoRI sites. sien.

So have to use other restriction enzymes to do the test...
some more the enzymes have different optimum temperature, so have to do sequential digestion and incubate twice..


Friday (1/8/08)
Analyze the product after restriction test n gel electrophoresis.
Funny... the band size wasn't what I wanted...
Went back to check all the gel photos...
I think I cut the wrong band on Monday...

Shit.. whole week's effort wasted.

Running experiments where results aren't guaranteed sure is a different story.
My lab notes and protocols

My Gel photos..

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