Master Mix Reagent Table.
|Ssofast Evagreen MM||10||180|
|Nuclease Free H2O||8||144|
1. Added each from greatest volume to least to make the master mix for each primer.
2. Pipetted 19 ul master mix into each well of a qPCR partial plate
3. Added 1 ul sample to each tube
|Sybr New Plate+Sybr cDNA 60 melt 2 Read|
|Initiation||95 C||10 min|
|Elongation||95 C||30 sec|
|55 C||1 min|
|72 C||30 sec|
|Repeat Elongation 39 times|
|Termination||95 C||1 min|
|55 C||1 sec|
|Melt Curve Manual ramp 0.2C per sec Read 0.5 C||55 - 95 C||30 sec|
|21 C||10 min|
Actin Melt Curve
The No template controls were clean and the amplification clear with good melt curves. Overall this looks really good. My next step is to test the new primers that came in today. I'll post tomorrow about what they look like.
You can see the raw qPCR data file here.