Bio-Rad
Sso Advanced Universal SYBR Green Supermix
172-5271
Info: Supplier Page
For these experiments we were assessing Aryl hydrocarbon Receptor activation in bone marrow derived dendritic cells. Precursor cells were isolated with Lympholyte M from whole bone marrow suspensions and grown in the presence of GM-CSF for 5-7 days. We then used flow cytometry and RT-PCR to assess changes in dendritic cells after AhR activation.
RT-PCR
cDNA reverse transcribed from whole cell RNA
For 10uL reaction, you need 5ul of SYBR Green Supermix, 250nm forward and reverse primers, and 100ng cDNA. This supreme requires a 30 second polymerase activation step, prior to denaturation, and annealing.
Perform cDNA titrations to see if 100g cDNA is optimal (we get cleaner results with 50ng)
We get reliable results with this SYBR Green Supermix, that can be stored for up to 1yr at -20C or three weeks at 4C.
If using a Bio Rad RT-PCR machine, you can fast load the cycle settings from their template list which is great!
Consistent results, and fast protocol
None
Bio Rad makes a really good SYBR Green Supermix.