One of the reviewers in my submitted paper ask me for the "verification of the reaction steps of PCR conditions". how to argument to this comment, do i have to add reference or i have to verify the steps again by performing PCR
Ghani Rehman
Assay verification and validation are essential when implementing a new diagnostic procedure. Analytical verification is defined as the process of establishing performance characteristics, such as the limit of detection, sensitivity, specificity, and reproducibility. Please have a look at this paper:
Article Analytical Verification of a PCR Assay for Identification of...
Good luck
Is this a paper about making an unusual form of pcr work? Does the result referred to in the paper rely on the PCR Not working.? Perhaps he is asking for positive and negative controls in your PCRs. It is not possible to say without more details
You have to cited the reference through which you replicate the PCR conditions.
Hi,
Ghani Rehman I think they need a citation for PCR amplification program you have used in the similar projects. Or maybe your knowledge about the time duration per step and cycles number are questioned.
Have you mentioned your PCR master mix content too? maybe asked about any modification such as adding Magnesium or concentration of DNA resource.
Even it would be asked about amplified probs, like size or concentration.
Good luck, Thanks.
Did you design a PCR-based assay? If so, you'll need to show a full set of controls. If you are using an established protocol, just cite the source (primers & cycling conditions).
Ghani Rehman
Assay verification and validation are essential when implementing a new diagnostic procedure. Analytical verification is defined as the process of establishing performance characteristics, such as the limit of detection, sensitivity, specificity, and reproducibility. Please have a look at this paper:
Article Analytical Verification of a PCR Assay for Identification of...
Good luck
Verification of PCR depends on your work as answered earlier but you can add reference (if you have followed the steps/conditions from other researchers), you may write about gradient PCR and you may also write about positive and negative control
You can add your reference or source if the protocol is already standardized by other researchers, if you did your own or modified it you can write your steps or evidence that it is really working , positive and negative controls should be included.
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