I am performing qRT-PCR for covid 19 using IC-VDE kit but there are two peak showed up in my positive control. The intensity also very low, below 1. Sometimes I found the sample also have ct between 35-36 almost near cut off for 37. Can someone give some explanation why?
Hello Rut Christine
1. This type of curve could result when the template concentration is high as there are chances of having non-specific background. So, you need to dilute your template.
2. Low intensity fluorescence could be because of the fluorophore. I would suggest you use another fluorophore having stronger fluorescent signal. Another possibility could be the system that you are using to perform qPCR. You may have to calibrate your system and have a look at the lamp. You may have to change the lamp. These options you can try later. Replacing the fluorophore with stronger fluorescent signal should be priority.
3. Do RNA quality check of your samples as well.
Good Luck.
