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Hi everyone,
Has anybody else actually counted the number of DNA capture beads going into the emPCR? I have found after numerous counts of Lib-L, and Lib-A and B capture beads that there can be a difference of at least 10% between what there should be, and is.
This won't make a difference to your final count (even though some blank capture beads may be being carried through the emPCR and giving false numbers of bound capture beads according to Roche), but it will affect the percentage of enrichment if calculated from a 'known' quantity of input capture beads according to the calculation in the protocol. Subsequently counts towards the top end of the range will actually be too high, affecting cpb interpretation and enrichment QC.
Has anybody else actually counted the number of DNA capture beads going into the emPCR? I have found after numerous counts of Lib-L, and Lib-A and B capture beads that there can be a difference of at least 10% between what there should be, and is.
This won't make a difference to your final count (even though some blank capture beads may be being carried through the emPCR and giving false numbers of bound capture beads according to Roche), but it will affect the percentage of enrichment if calculated from a 'known' quantity of input capture beads according to the calculation in the protocol. Subsequently counts towards the top end of the range will actually be too high, affecting cpb interpretation and enrichment QC.
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