Earlier I got amplification, but now not getting amlification with the same protocol?

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Katie A S Burnette

My guess is that your primers have degraded. Make a fresh batch & I'm guessing it will work again. Also, where is your positive control?

Other possibility is that you have too much DNA (or too many PCR-inhibiting molecules) in the DNA extractions. Dilute the DNA 1:10 and see if that helps.

Good luck!

Deepti Prabha

Thank you. I appreciate your suggestions and will make an effort to implement them.