Spidermonkey's Sporadic Lab Stuff
by Spidermonkey
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Hello World from the lab!
Part spider - part lab monkey, will do science for cash, strawberries, or for a place to be that's out of the rain (but preferably for cash).
The rest of what I get up to goes into the Tunnel of Goats :)
Urgent Science Stuff:
Please help reform English libel law
Current UK libel laws are very bad for science and free speech.
Cool science stuff:
RCSB Protein data bank: Molecule of the Month in alphabetical order
The PCR Song It's the little things that keep you going...
NCBI's Entrez Gene My favourite starting point for finding out what is known about any gene of interest.
NCBI's PubMed Where to go to find pretty much all research published in the last 30 years or so (may go back a lot further now). All newly published research is quickly added to the site.
The Genetic Code - table of the DNA/RNA triplet codes for amino acids. This is how DNA codes for protein.
Another PCR song Disco frenzy :)
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from yesterday and today.
9th Dec 2009, 00:12
comments (2)
Light from the luciferase reporter transfected into the cells. THis assay worked really nicely. More than can be said for my cloning, but I do have some colonies this time... Will find out tomorrow if any of them contain my special bit of DNA. I'm not holding my breath.
7th Dec 2009, 01:10
comments (0)
Am still trying to get my cloning to work (this time using maximum superstition), but luckily I have other work to do that has usually runs along more successful lines, like this one here. 1 flask HuH7 cells transfected with plasmids for nuclear receptor gene of interest, firefly luciferase reporter and green fluorescent protein (to check that transfection has worked). Here I see green cells in good enough numbers to do the next bit of the assay (plate them out into a 96-well plate and treat them with different compounds of interest). Tomorrow I should be able to harvest the plate and see which compounds interact with my favourite protein of the day. It will give me numbers, which I can plot and subject to basic statistical testing. This is comforting.
4th Dec 2009, 22:21
comments (2)
After 3 days there is still no growth (usually leaving them overnight is more than enough). There is not much hope left for these little bugs. Trying again. again.
3rd Dec 2009, 22:40
comments (0)
More agar for plate making and finding the right sequence to amplify up for my next attempts at cloning. Sooner or later something has to work. If an infinite number of Spidermonkeys were given typewriters etc. etc.
2nd Dec 2009, 21:55
comments (4)
This was from yesterday. 1 flask of cells transfected with nuclear receptor and luciferase reported plasmids + green fluorescent protein as a marker for transfection having taken place (i.e. if the cells have taken up the one that makes them turn green, there's a good chance they've taken up the other plasmids too). Checking for green fluorscence uses the pretty blue light.
Forgot to take a picture of the actual cells though. They were nice and green too. Next time, if there's nobody about :)
2nd Dec 2009, 21:42
comments (0)
The colonies are very tiny and slow growing, but hopefully they still contain the plasmid (so they will make my bit of DNA for me). Have bunged some into culture over night (in the broth from the bottom pic)It was impossible to pick single colonies as they are so tiny and so slow growing so I've just taken several dabs of a couple of the stripes of bugs and am hoping some of them will grow up overnight in liquid culture. Then I can steak some of that out onto new plates and pick single colonies later. In the meantime I can try to isolate the DNA from these and then (if I get enough) I can see if it will work with the mammalian cells (middle pic). In other words I'll be able to actually do an experiment!
Will see how things look in the morning.
30th Nov 2009, 23:56
comments (8)
from yesterday morning.
29th Nov 2009, 22:10
comments (3)
