What is the typical yield I should get from my library prep?
The typical library concentration you can expect from Lexogen libraries amplified with an optimal number of PCR cycles is ~1 - 3.5 ng/µl1.
This amount of library is sufficient for pooling and sequencing.
Lower library yields can also be sufficient for pooling and sequencing. To check whether your library amounts are sufficient, use our Library Quantification Calculation File. Please also check with your sequencing provider lanemix concentration and volume requirements.
Lanemixes that are too diluted after samples pooling can be concentrated into a smaller volume using the Lexogen Purification Module with Magnetic Beads (Cat. No. 022), using 0.9 - 1x Purification Beads (PB) and eluting the re-purified pool in a smaller volume of Elution Buffer (EB).
If your library yield or concentration is still not sufficient for sequencing, you can reamplify your libraries or lanemix using the PCR Add-on and Reamplification Kit (Cat. No. 208).
Please check these FAQs for library reamplification kit options and compatibility details:
These typical library concentration ranges do not apply to the miRVEL Discovery Small RNA-Seq Library Prep (Cat. No. 242). For this kit, there is no defined library concentration threshold that indicates over-amplification. With intact, high-quality RNA inputs, the standard protocol should yield optimal libraries. Over-amplification is instead identified by electropherogram profiles showing characteristic rolling humps at approximately 400 bp, 500 bp, or 900 bp.