On real time PCR, cDNA kits etc.

Hi there

A couple of (good) news for people dealing with real time PCR:

1-The Mj machine at the SBM can read a signal from a 8ul reaction. That means you can set your reaction using only 4ul of SybrGreen/replicate. However you really need a good pipette since small pipetting errors become critical at this point.

2- The QuantiTect cDNA kit from Quiagen is so far the least expensive and more efficient kit I had the chance to try. There is only one caveat: you need RNA already digested with DNAse. The kit includes a "genomic DNA wipeout reagent" to take care of genomic DNA before the cDNA synthesis, but as far as I can tell it's useless. Scaling don the reaction works fine, the kit works like a charm and it is really a pleasure to use (it's a 2 step process that only takes about 30').

Hope this is helpful

Till next one
Gabriele Amore

About Microarrays

Dear all,

since I have the feeling that the interest in microarray , gene function annotation and experimental next steps is increasing in the Institute I would like to initiate something like a round table, even very relaxed. We can maybe meet outside this walls where we can not only enjoy discussions on scientific things but also a cold beer and some crackers. So whoever is interested can contact me via this blog or directly via mail: aluedeking@meeresforschung.de

So, lets start something

Alexander Luedeking

Quantitative PCR with in just 10 ul!!

Hey there
Are you still running qPCR reactions in 25 ul?? Really?? well here's the scoop: you can do it in just 10ul and it works just as fine. I did it 7 times (that is 7 full 96 well MJ white plates) and it went just allright.
For further details call me at 315

sincerely yours

Gabriele Amore