...
How can I prevent overcycling?
Perform the qPCR assay to determine the optimal number of PCR cycles required for library generation.
For similar samples , reduce the number of PCR cycles for library amplification (Endpoint PCR) to prevent overcyclingover-cycling.
You can estimate the number of cycles to reduce according to the yield of overcycled libraries. To achieve half the yield, subtract 1 PCR cycle. To achieve 4x less yield, use 2 fewer cycles. To achieve 10x less yield, use 3 fewer PCR cycles.