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:
- Fast and simple protocol
- Hands-on time is around 10 minutes
- The total protocol time is only 1.5 hours
- Easy procedure based on:
- Ready-to-use master mix for easy setup of reactions
- Less reaction components to simplify bench work
- Less magnetic beads used for cleanup procedures: This can reduce more than 50% of the beads consumption
- Guaranteed high library conversion efficiency
- Low input cell-free DNA: From 1 ng to 50 ng
Comparison of library conversion efficiency with different samples under the same condition.
Comparison of library yield with different samples under the same condition.
