Bitan:qPCR: Difference between revisions
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=qPCR recipe= | =qPCR recipe= | ||
Total volume per well is 25 μL.<br> | Total volume per well is 25 μL.<br> | ||
iQ SYBR Green Supermix | 12.5 μL iQ SYBR Green Supermix<br> | ||
100-500 nM each primer<br> | |||
1 μL sample<br> | |||
ddH<sub>2</sub>O to 25 μL<br> | ddH<sub>2</sub>O to 25 μL<br> | ||
Revision as of 13:41, 8 December 2009
Quantitative PCR uses fluorescent dye to quantify the relative difference in sequence copy number between two samples. The dye used is SYBR Green which binds to dsDNA but not to ssDNA, when comparing two qPCR reactions of a given sequence SYBR Green's signal is proportional to sequence copy number.
The Bitan lab does not have a qPCR machine. There is one in the de Vellis lab on the third floor that you can use if you get permission from Cristina Ghiani (cghiani at mednet stop ucla stop edu). You must also sign up to use the machine at least a day in advance of your experiment using the calendar that hangs over the machine.
Bio-Rad iQ SYBR Green Supermix Manual
The manual for this kit is here.
qPCR recipe
Total volume per well is 25 μL.
12.5 μL iQ SYBR Green Supermix
100-500 nM each primer
1 μL sample
ddH2O to 25 μL
