Rob™ qPCR set-up reproducibility for 1μl template

The template is a single stranded DNA fragment containing the beta-actin amplicon sequence. Primers are specific for mouse beta-actin. 

Result:
CqAverage: 21.35
Standard deviation (Cq): 0.08
CqMax – CqMin: 0.30

Run conditions:
QPCR: HotStart activation, 2 min at 95℃ followed by 40 cycles with 10 s at 95℃ and 10 s at 60℃.
Run time: 45 min.
Set-up: 18 μl master mix was dispensed using multi dispense followed by 1 μl template using single dispense from one tube. Run time: 8 min 14s.

Reagents:
qPCRBIO SyGreen Mix from PCRBiosystems was selected because it is the fastest and most sensitive kit available on the market. It combines the latest polymerase and buffer technology with a proprietary intercalating dye that doesn’t inhibit PCR.

QPCR instrument:
Mic qPCR-instrument from Bio Molecular Systems is the most precise qPCR instrument available today. Virtually no temperature differences between samples during the run combined with the magnetic induction heating ensures highest possible quality on data.
Reproducibility of Mic is within Cq 0.20.

Conclusion:
The precise liquid handling performance of Rob™ allows precise QPCR analysis for template volumes down to 1 μl. The reproducibility data on 1 μl is better than the reproducibility of 96/384 plate qPCR instruments.

Download this information, click Performance note: qPCR set-up reproducibility for 1µL.