![]() But how much water do you need to add? The amount of PCR-grade water to add depends on the number of nanomoles (nmoles) for that oligo primer. Next, you need to reconstitute your primers with PCR-grade water to make a 100 μM stock solution. Otherwise, if the powder is stuck in the lid while opening the tube, you could potentially damage your primers. Reconstitute your stock primersįirst things first, you should briefly (approximately 30 seconds) centrifuge your primers to pull all of the lyophilised powder to the bottom of the tube. To prepare primers for PCR, just follow these two simple steps: 1. Use filter pipette tips to prevent contamination via pipetting.Use PCR-grade water (DNase- and RNase-free) to reconstitute and dilute your primers.This is to avoid contamination of your stock and working primer solutions. Make sure you work in a DNA-free environment.But how do you prepare lyophilised (freeze-dried powder form) oligo primers for use in PCR? If you are unable to prepare the primers straight away, then store them at 4 oC until the time comes.īefore you start, here are some general tips when working with PCR primers: So you have designed and ordered your primers and now they have finally arrived. >Use code 20QPCR to get 20% off<< Before you start ![]() Mastering qPCRĪ video tutorial on how to dilute new qPCR primers can be found in our Mastering qPCR course In this article, I will explain how to easily prepare PCR stock primers and how to dilute them into a working primer solution.
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