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Old 08-15-2019, 01:49 PM   #1
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Location: Philadelphia

Join Date: Aug 2019
Posts: 3
Default Pre-Hybridization PCR not running efficiently

Hello all,
I am working on building my libraries for an NGS project. I am running into a problem in which my pre-amplified libraries have concentrations ranging from 8 ng/uL up to 15 ng/uL. However, when I attempt to amplify these libraries using a Phusion master mix and the primers that are compatible with our adapters for a 12 cycle PCR, the product qubits much lower (.5 ng/uL - 1 ng/uL) and appears to have not amplified. Running a normal QC PCR however I see product on my PCR gel indicating that the adapters ligated properly and are working with the primers. If anyone has any idea what my be causing this or possible troubleshooting steps that would be much appreciated!
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