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The number of cycles for library amplification depends on the amount of input dsDNA as well as your downstream application.
For PCR-amplified libraries, such as those used in whole genome sequencing (WGS) applications, the minimum recommended number of PCR cycles for different dsDNA input amounts is outlined below (expected yield is ~4 nM final library):
Input of dsDNA (ng) | Minimum PCR cycles |
≥25 | 4 |
10 | 6 |
1 | 9 |
If the constructed libraries will be subsequently used for hybridization enrichment, the minimum recommended number of PCR cycles for different dsDNA input amounts is outlined below (expected yield is ≥500 ng of material for capture):
Input of dsDNA (ng) | Minimum PCR cycles |
250 | 4–5 |
100 | 6–7 |
25 | 8–9 |
10 | 9–10 |
1 | 13–14 |
Note that library amplification may not be required for some applications. However, if library amplification is not performed, full-length adapters must be used for adapter ligation.
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