Friday, June 27, 2014

How to elute delivered DNA on Whatman paper

Usually,  we deliver DNA (1 ug) dried on a Whatman paper and the spot is circled with a pencil to mark the spot. That is the most convenient way to post/mail DNA abroad. It's better and more cost-saving than shipping DNA solution in a tube.
Once the DNA reaches the lab,  we would cut out the spot demarcated by pencil and place it in a purification column and elute it with TE buffer.  However,  a purification column costs sgd 1.70 per item (or lesser now).
Another cheaper way to elute the DNA is by using a 0.5 ml tube (with the cap removed) and stack it into a 1.5 ml tube.  Then by means of a syringe,  the bottom of the 0.5 ml tube is perforated by repeated jabbing. Next,  the cut-out DNA on paper is added into the 0.5 ml tube and sufficient TE buffer (e.g. 30 ml) is added to soak the cut-out piece. The assembly is the spun at max speed for 1 to 2 min to elute the DNA solution into the 1.5 ml tube. See figure for reference.

Thursday, June 19, 2014

Mnemonic - RNA pol II holoenzyme (my ref)

There are total of six general transcription factors (complexes):

TFIIA (Assist) TFIID to (Dock) to DNA. TFIIB (Binds) to TFIID to recruit RNA pol II. TFIIF (Fidelity) bound to RNA pol II prevents unspecific docking. TFIIE (Elongation) and TFIIH (Helicase) are recruited to docked RNA pol II and prepare from initiation to elongation phase.


Friday, June 13, 2014

"Good Samaritan corner" in the lab

Good Samaritan corner is needed in research lab. In my lab, common reagents/solutions were prepared by a single person and shared by many people. That took a toll on that person (especially when someone later complained about lack of solutions that affected experiments) and since then, there was no longer common reagents/solutions and everyone was supposed to prepare his/her own solutions.

I'm planning to assign a corner (or other places) in the lab "Good Samaritan corner" where we can volunteer to prepare solution without any expectation of its availability. If the solution is unavailable, no one should should, "solution A is empty!". The corner will be "you reap what you sow", in which the availability of common solutions is a shared volunteerism.