The NGS Cell Free DNA Library Prep Kit (illumina and MGI Platforms) was developed for the construction of high quality cell-free DNA (cfDNA) libraries using 1 ng to 50 ng of cell-free DNA as input. The kit has a simple work flow and a fast procedure. Multiplexing of the cell free DNA library is possible based on the index type.
NGS Cell Free DNA Library Prep Kit Workflow
The main source of cell-free DNA (cfDNA) is derived from apoptotic hematopoietic cells in blood and found in the plasma. The length of the cfDNA is about 150-200 bp in length. Circulating tumor DNA (ctDNA) derived from malignant tumors is a part of cfDNA. Both cfDNA and ctDNA can be used as a noninvasive biomarker since it offers a better approach in comparison to invasive tissue biopsies.
NGS has been used for cfDNA and ctDNA sequencing in the field of liquid biopsy as it provides a whole genome level of molecular profiling. One of the hurdles for cfDNA sequencing is the difficulty of library preparation from the limited amount of cfDNA obtained from plasma. Our cell-free NGS kit makes it easy to get enough libraries from limited input in just 1.5 hours.
Three index types are available for the NGS Cell Free DNA Library Prep Kit of the illumina platform:
Non-index (Cat.# 30029): Libraries do not have index.
Index(Cat.# 30031): Each of our index primers contains a unique 6-base index sequence that can be used for sample identification. Total 48 library multiplexing is possible. Index information can be downloaded here.
Unique dual index (Cat.# 30033): Cell-free DNA library multiplexing up to 96 samples is possible with the unique dual indexes. We have developed a Four-Base Difference Index System. The system have at least 4 bases different from each other in the 8 bases index length. The primers effectively minimize sequencing errors such as mis-assignment, index hopping, index contamination etc. Index information can be downloaded here.
Indexes are available for the MGI platform kits (Cat.# 34031).
Kit advantages:
Comparison of library conversion efficiency with different samples under the same condition.
Comparison of library yield with different samples under the same condition.
The NGS Cell Free DNA Library Prep Kit (illumina and MGI Platforms) was developed for the construction of high quality cell-free DNA (cfDNA) libraries using 1 ng to 50 ng of cell-free DNA as input. The kit has a simple work flow and a fast procedure. Multiplexing of the cell free DNA library is possible based on the index type.
Norgen’s NGS Library Quantification Kit (for Small RNA-Seq) offers a PCR-based detection procedure to quantify NGS libraries (specifically Small RNA-Seq) of a wide spectrum of concentrations. The kit consists of a specially designed primer mix compatible with the Illumina system, that is used in conjunction with the provided 2X Real-Time PCR Master Mix to amplify a library of unknown concentration. The unknown library is accurately quantified by using a standard curve constructed from the provided DNA Standard (range from 20 pM to 2 fM) on a Real-Time PCR System. The kit is specially optimized to quantify Small RNA-Seq libraries with DNA standards that have similar size to a Small RNA-Seq library. However, it could also be used with other types of NGS libraries.
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Storage Conditions
Upon receipt, store Norgen’s NGS Library Quantification Kit (for Small RNA-Seq) at -20°C or lower. Avoid multiple freeze-thaw cycles. If needed, prepare smaller working aliquots and store at -20°C or lower.
| Component | Cat. 61600 (100 reactions) |
|---|---|
| 2X Real-Time PCR Master Mix | 1 mL |
| NGS Library Quantification Primer Set Mix | 600 µL |
| Quantified NGS Library Standards (for Small-RNA Seq)* | 5 x 80 µL Standards |
| Product Insert | 1 |
* Quantified Library Standards are provided as 20 pM, 2 pM, 200 fM, 20 fM and 2 fM
Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.
The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit. Details of the target and priming specificity are included in the individual handbooks above.
Packaged, optimised and ready to use. Expect Better Data.
Primer and probe mix (150 reactions)
Reverse Transcription, target specific primers (RNA genome viruses only)
Copy number standard curve (sufficient for multiple standard curves)
Internal extraction control – Read through VIC channel*
Endogenous control (150 tests)
RNAse/DNAse free water
*alternative fluorophores available on request