5.6 qPCR | ISB Server Wahoo

Submitted by kdorfman on Fri, 04/19/2013 - 17:05

Q-PCR after RT

Housekeeping gene:

Gene At5g25760 (Ubiquitin Congugating enzyme 21) UBC21

Forward primer TCTCTCTCTCTCTCTCTCGCTCTC Reverse Primer TGATGCCTGCATCTCTAATTTCCC

Re-think the need to have exactly the same amount of RNA in each cDNA reaction.

Set up reactions in ISB. Use Qiagen 204054

Load reactions and take to Sam's lab.

Eppendorf Realplex2 Master Cycler

Reaction set up

component	µL/rxn	aliquot for 16 rxns
2x quantifast SYBR Green PCR master mix	10	176
qPCR forward primer 10 µM	1	32
qPCR reverse primer 10 µM	1	32
Template cDNA	1	-
RNAse free water	7	(use H2O from RT)
------------------------------------------------	----	-----
Total	20