The PCR-free NGS DNA Library Prep Kit was developed for construction of DNA libraries for next generation sequencing (illumina platform). The kit adds 3′-dT-tailed library adapters to both ends of DNA fragments efficiently. The kit uses double strand DNA fragments (blunt and/or sticky) as input DNA for NGS library construction, and is compatible with DNA fragments generated from both enzymatic methods (BioDynami DNA fragmentation enzymes etc.) and physical methods (sonication, nebulization etc.).
PCR-Free NGS DNA Library Prep Kit Workflow
PCR amplification is a standard step for library preparation of Next-Generation Sequencing. The PCR is used to amplify fully ligated DNA fragments and to add index information to the libraries. The indexing is for the pooling of the library samples in an effort to reduce the sequencing cost.
However, PCR introduces uneven amplification of DNA in some DNA regions with extreme GC-contents and secondary structures. This bias can cause very low sequencing coverage in such regions. DNA sequencing in these regions is still a huge challenge.
PCR-free library prep can reduce library bias and minimize sequencing gaps. The sequencing data from PCR-free library samples have even genomic coverage with few gaps and better depth in GC-rich regions. Our PCR-free NGS kit offers optimal coverage in the regions that are traditionally difficult, such as high-GC content regions, low-GC content regions, and repetitive sequence regions.
Two index types are available for the kit:
Non-index: Libraries do not have index.
Index: Each library contains one i5 index and one i7 index. Library multiplexing up to 96 samples is possible. List of indexes can be downloaded Here.
Kit advantages:
The PCR-free NGS DNA Library Prep Kit was developed for construction of DNA libraries for next generation sequencing (illumina platform). The kit adds 3′-dT-tailed library adapters to both ends of DNA fragments efficiently. The kit uses double strand DNA fragments (blunt and/or sticky) as input DNA for NGS library construction, and is compatible with DNA fragments generated from both enzymatic methods (BioDynami DNA fragmentation enzymes etc.) and physical methods (sonication, nebulization etc.).
Volcano3G® RT-PCR Probe 2x Master Mix contains all components required for general RT-qPCR
– optimized reaction mix for sensitive and reliable results
– engineered, truly thermostable Taq DNA polymerases with reverse transcriptase activity
– fast start function due to a hotstart aptamer formulation which prevents unspecific amplification at lower temperatures (<57°C)
– Pipetting aid in form of a blue dye for better visibility during aliquoting into well-plates
– included lysis function*
* = A sample extraction and sample lysis is not necessary as immediate hot PCR cycling conditions can brake cell- and virus membranes. Please see our published references to this product.
Note: Volcano3G® RT-PCR Probe 2x Master Mix is a unique master mix. If you are using it for the first time, we recommend to design primers to have higher melting/annealing temperatures (above 65°C). Additionally, for assay setup it is advised to perform RT-PCR test reactions with temperature gradients (i.e., for the annealing and extension steps) to identify the most proficient PCR protocol for your needs using our Volcano DNA polymerase products.
For research use and further manufacturing.
In case you are aiming to use our RUO products as components or for your development of e.g. an IVD medical device, please contact us.
Volcano3G® RT-PCR Probe 2x Master Mix contains all components required for general RT-qPCR
– optimized reaction mix for sensitive and reliable results
– engineered, truly thermostable Taq DNA polymerases with reverse transcriptase activity
– fast start function due to a hotstart aptamer formulation which prevents unspecific amplification at lower temperatures (
This kit provides a fast and simple spin column procedure for isolating genomic DNA from saliva samples collected and preserved using Norgen’s Saliva DNA Collection and Preservation Devices (Cat. 49000), as well as fresh saliva samples.
Saliva DNA purified using Norgen’s Saliva DNA Isolation Kit Dx kit is of the highest quality, and can be used with any downstream in vitro diagnostic application employing enzymatic amplification or other enzymatic modifications of DNA followed by signal detection or amplification.
Background
Saliva represents an excellent non-invasive alternative to blood collection. Human genomic DNA extracted from buccal epithelial cells and white blood cells found in saliva can be used in various applications in diagnostics. Saliva DNA can be used for the detection of biomarkers to diagnose a disease, follow the diseases progress or monitor the effects of a particular treatment. Saliva DNA can also be used to diagnose particular types of infections. Isolation of DNA from saliva has become an attractive alternative to isolation from blood or tissue due to the fact that sample collection is non-invasive, the samples can be collected by individuals with little training, and no special equipment is required. Norgen’s Saliva DNA Isolation Kit provides a fast and simple procedure for isolating genomic DNA from both preserved saliva samples and fresh saliva samples.
NOTE: This product is not available for sale in the United States.
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| Kit Specifications | |
| Maximum Saliva Input | 0.5 mL preserved saliva 0.25 mL fresh saliva |
| Average Yield from 0.25 mL of Saliva* | 3 – 7 μg |
| Average Purity (OD260/280) | 1.7 – 2.1 |
| Time to Complete 10 Purifications | 30 minutes |
* Average yield will depending on the donor
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. All solutions and plastics can be used until the expiration date specified on their labels. The Proteinase K can be stored at either room temperature or 4oC.
| Component | Cat. Dx45400 (50 preps) |
|---|---|
| Lysis Solution | 30 mL |
| Proteinase K in Storage Buffer | 1.2 mL |
| Binding Solution | 12 mL |
| Wash Solution | 18 mL |
| Elution Buffer | 12 mL |
| Spin Columns | 50 |
| Collection Tubes | 50 |
| Elution Tubes (1.7 mL) | 50 |
| Product Insert | 1 |