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.
West Nile Virus (WNV) belongs to the RNA virus family of Flaviviridae. It is known to spread primarily through arthropod vectors such as mosquitoes. The virus infects mainly birds but is also reported to infect humans as well as pets such as cats and dogs. In humans, the majority of the infections caused by WNV are asymptomatic. However, some individuals (the majority of the confirmed cases) could enter a second, febrile stage with flu-like symptoms – commonly known as West Nile Fever. In a more serious stage, the disease becomes neuroinvasive, causing meningitis or encephalitis. Such severe conditions could lead to mortality. As the symptoms of WNV are very similar to other common diseases but can be fatal at a severe stage, it is important to distinguish the virus early in the diagnosis, particularly at the molecular level.
WNV TaqMan RT-PCR Kit, 100 reactions
WNV TaqMan RT-PCR Probe/Primer Set and Controls, 100 reactions
Figure 1 / 3
Click for expanded view
Storage Conditions and Product Stability
All kit components can be stored for 1 year after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival.
| Component | Cat. TM44250 (100 preps) | Cat. TM44210 (100 preps) |
|---|---|---|
| MDx TaqMan 2X PCR Master Mix | 2 x 700 μL | – |
| WNV Primer & Probe Mix | 280 μL | 280 μL |
| WNV Positive Control | 150 μL | 150 μL |
| Nuclease-Free Water (Negative Control) | 1.25 mL | 1.25 mL |
| Product Insert | 1 | 1 |
Norgen’s Plasma/Serum Circulating DNA Purification Kits (Slurry Format) provide efficient methods for the purification of fragmented free-circulating DNA from human plasma or serum. The kit is able to isolate all sizes of circulating DNA. The slurry format provides an advantage over other available kits in that it does not require extension tubes for the purification of free-circulating DNA from large sample volumes. DNA can be isolated from either fresh or frozen samples using this kit. The kit will also isolate viral and bacterial DNA from plasma/serum. Typical yields of purified free-circulating DNA will vary depending on the input sample (1-100ng/mL circulating DNA in human plasma), with more concentrated samples tending to yield more free-circulating nucleic acids. The purified, inhibitor-free plasma/serum free-circulating DNA is eluted in an elution solution that is compatible with PCR, qPCR, methylation-sensitive PCR and Southern Blot analysis.
Plasma/Serum Circulating DNA Purification Mini Kit (Slurry Format)
Norgen’s Plasma/Serum Circulating DNA Purification Mini Kit (Slurry Format) provides a fast, reliable and simple procedure for isolating circulating DNA from various amounts of plasma/serum ranging from 50 μL to 400 μL. Preparation time for a single sample is less than 30 minutes.
Plasma/Serum Circulating DNA Purification Midi Kit (Slurry Format)
Norgen’s Plasma/Serum Circulating DNA Purification Midi Kit (Slurry Format) provides a fast, reliable and simple procedure for isolating circulating DNA from various amounts of plasma/serum ranging from 400 μL to 2 mL. Preparation time for a single sample is less than 45 minutes.
Plasma/Serum Circulating DNA Purification Maxi Kit (Slurry Format)
Norgen’s Plasma/Serum Circulating DNA Purification Maxi Kit (Slurry Format) provides a fast, reliable and simple procedure for isolating circulating DNA from various amounts of plasma/serum ranging from 2 mL to 10 mL. Preparation time for a single sample is less than 45 minutes.
Figure 1 / 10
Click for expanded view
| Kit Specifications | |
| Minimum Plasma/Serum Input | 2 mL |
| Maximum Plasma/Serum Input | 10 mL |
| Size of DNA Purified | ≥ 50 bp |
| Elution Volume | 100 μL |
| Time to Complete 10 Purifications | 15-20 minutes |
| Average Yields | Variable depending on specimen |
Note: Do not exceed the recommended sample input volume of 10 mL.
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
| Component | Cat. 50600 (50 preps) | Cat. 51200 (20 preps) | Cat. 51300 (10 preps) |
|---|---|---|---|
| Lysis Buffer D | 65 mL | 125 mL | 3 x 125 mL |
| Slurry A1 | – | 6 mL | – |
| Slurry A2 | 12 mL | – | 12 mL |
| Binding Buffer B | 20 mL | 6 mL | 20 mL |
| Binding Buffer C | – | – | 30 mL |
| Wash Solution A | 2 x 38 mL | 2 x 38 mL | 3 x 38 mL |
| Elution Buffer B | 8 mL | 8 mL | 15 mL |
| Mini Filter Spin Columns | 50 | 20 | – |
| Midi Filter Spin Columns | – | – | 10 |
| Midi Collection and Elution Tubes | – | – | 20 |
| Collection Tubes | 50 | 40 | 10 |
| Spin Columns | – | 20 | 10 |
| Elution Tubes (1.7 mL) | 50 | 40 | 10 |
| Product Insert | 1 | 1 | 1 |