I am using an automation nucleic acid extraction system to extract HBV DNA from sera and used an externally added internal control along with it during extraction. In the automation nucleic acid extraction using magnetic beads, I have increased the beads concentration and obtained a good early Ct result, when tested on RT PCR, for the virus gene but the internal control added to it was showing late Ct. This is a singleplex reaction and which parameter do I have to alter to get an early Ct for the internal control too?
Please any one answer me?
Dear Umadevi Subramanian
These links might be useful have a look:
1. https://www.mdpi.com/2075-4418/11/6/1091/pdf
2. https://www.scielo.br/j/rimtsp/a/hbQtF546GT5MxyGTsXkBLjC/?format=pdf&lang=en
3. https://assets.publishing.service.gov.uk/government/uploads/system/uploads/attachment_data/file/926410/Understanding_Cycle_Threshold__Ct__in_SARS-CoV-2_RT-PCR_.pdf
4. https://virologyj.biomedcentral.com/articles/10.1186/s12985-021-01559-3
5. https://www.sciencedirect.com/science/article/abs/pii/S0379073821000955?dgcid=rss_sd_all
Kind Regards
Qamar Ul Islam
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