NGS Library Quantification Kit

Simple to use kit for checking your NGS library quantity and quality before you sequence it

The YouSeq NGS Library quantification kit employs qPCR as a highly specific and Sensitive method to calculate the amount of DNA library present with successfully incorporated adapters. This kit will help improve library loading concentration and therefore, improve sequencing result for each patient so reducing risk of patient repeats.
qPCR is the only technically capable of a library quantification of total sequencable product. Other technologies will include non-specific products in the quantification meaning reduction in quantification accuracy.

Results are compared to a set of calibrated standards for precise quantification.

Dilution buffer is also supplied to dilute your DNA library accordingly, ready to be sequenced.

Library Quantification Handbook

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96 rxns (Illumina) $128.70

Cat no. YS-LQC-IL-96

96 rxns (Illumina) $128.70

Cat no. YS-LQC-IL-96

384 rxns (Illumina) $422.50

Cat no. YS-LQC-IL-384

960 rxns (Illumina) $1,007.50

Cat no. YS-LQC-IL-960

96 rxns (IonTorrent) $128.70

Cat no. YS-LQC-IT-96

384 rxns (IonTorrent) $0.00

Cat no. YS-LQC-IT-384

960 rxns (IonTorrent) $0.00

Cat no. YS-LQC-IT-960

Pack size:

Available for direct shipping to United States.

Kit contents

- Library specific qPCR primers
- qPCR MasterMix premixed with SYBRgreen
- 4 x positive control templates to create standard curve
- NGS library dilution buffer

Application

Specifically quantifies sequence ready library only:

- Precise across a very broad dynamic range
- Ideal for automated liquid handling
- Less expensive than alternative methods
- Highly sensitive- accurate quantification of very dilute libraries
- Consumes only small amounts of sample

Technical specification

qPCR is the most accurate method for quantification of an NGS Library. It is also far more cost effective and requires less bench time, especially in higher throughput laboratories vs Bioanalyzer of Spectrophotometer

Simply compare your library to the standard curve for precise quantitation

Your Questions Answered

Why is qPCR better than standard methods for NGS library quantification?

There are disadvantages to nearly all other methods for library quantification. Typical alternative methods include Qubit® (spectrophotometry) or Bioanalyser®/Tapestation® (electrophoresis). Neither of these methods are for suited to high-throughput of samples, requiring laborious and error-prone manual liquid handling. Furthermore they both measure total nucleic acid concentration which is not an accurate measure of sequence-ready library. Our qPCR kit is highly specific in identifying sequence-ready library only. In addition both of these methods lack sensitivity compared to qPCR and are typically more expensive.

What causes variability in qPCR Library quantitation data?

Inaccurate liquid handling is the most common reason for variability. We are happy to give guidance on good pipetting technique and good laboratory practice. Even simple things like ensuring that all standards are thoroughly vortexed to ensure they are correctly resuspended will help to ensure good data quality.

What are the storage recommendations for YouSeq NGS Library Quantification Kits?

Upon receipt, store the kit at -20°C. When stored and handled correctly, the kit components will remain perfect for at least six months from the date of receipt. All components of the kit – are stable for more than 20 freeze/thaw cycles.

Why does the NGS Library QC only have a 4 point standard curve?

Using 4 points instead of (for example) 6 saves previous wells on every plate that you run. The primers in the kit are highly efficient ~100%. We have demonstrated (during our kit development process) that the kit works over a wide dynamic range. Therefore, it is fair to extrapolate the 4 point standard curve in both directions and still deliver highly accurate results for samples that may fall out side of the range of the standard curve.