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What are the possible reasons for low pre-library yield? How to solve it?

View: 63 / Time: 2026-03-25
Possible Reason Solution
Input amount of nucleic acid is too low

If the sample is a co-extracted total nucleic acid, quantification of RNA and DNA should be performed separately using Qubit, and library amplification should be carried out according to the recommended cycles in the manual.


During the experiment, RNase-free consumables must be used, and the operating environment should be free of RNase contamination, as this can affect the efficiency of RNA reverse transcription.


If the nucleic acid sample contains excessive amounts of chelating agents, guanidine salts, phenol, proteins, ethanol, or other impurities, it can impact the activity of RNA reverse transcriptase and the efficiency of DNA enzymatic digestion. It is recommended refer to Step 7: Post-amplification Clean-up to purify the nucleic acid samples using 2X NadPrep SP Beads, replace the elution reagent with Nuclease Free Water, and then proceed with library preparation.
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