Hellow
I am performing illumina mini seq of 18 TB targeted gene. So when i amplified my targeted gene,i have to add internal control with master mix. But i am not clear about the appropiate function of this IC in PCR . Moreover for sequencing, I have to give individual internal control ,positive control and Negative control with my sample.So what will be the function of IC in sequencing??
Hello Fahim Alam Nobel
The use of an internal control (IC) is necessary to differentiate the true negative results from those consequents from a failure in some step of the nucleic acid test. The main purpose of IC is to maximize test sensitivity by identifying inhibitory (i.e., non-amplifiable) specimens that have the potential to generate false-negative results.
Incorporating an IC into PCR-based tests will increase sensitivity by enabling the user to identify and retest those samples inhibitory to PCR. A positive IC result indicates that amplification has occurred and thus gives you an assurance that the negative test results are truly negative.
Sample-to-sample variation, localization of infection, low target concentration are among other factors that could cause false negative results. So, it is necessary that you introduce IC into each test sample that is coamplified with the target nucleic acid to prove the functionality of the reaction mix for amplification of the target. This combination rules out inhibition, among other malfunctions, and confirms that a negative result is truly negative.
Best.
Thank you so much. I would like to know the function of IC in sequencing.As we created library pool for sequencing ,we add positive control,negative control and internal control.So here what is the function of IC?
