Novallele™ Assay Amplification Reaction Using NEXTGENPCR

Dana Sullivan, Product Manager at Canon BioMedical on September 17, 2018

NEXTGENPCR has been shown to speed the PCR amplification of DNA. In this technical note, we use NEXTGENPCR in the amplification step of our Novallele assay for the G6PC gene. Using the advanced mode function of NEXTGENPCR, we were able to reduce the time for a 35 cycle amplification to only 4 minutes and 30 seconds.

NEXTGENPCR amplification profile (Advanced Mode)

Cycles Temp Time
1 95°C 30 sec
10 95°C
60°C
72°C
2 sec
5 sec
2 sec
25 95°C
60°C
72°C
1 sec
3 sec
1 sec
1 72°C 10 sec


We subsequently analyzed the product by agarose gel electrophoresis to verify the length and look at the yield of the amplification product. We then analyzed by high-resolution melting analysis and determined the results were as expected for the Novallele assay.

 

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