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:

• • • Total time: 1 hr
    • Hands-on time: 5 min
    • Easy procedure: Ready-to-use master mix & Less reaction components
    • Input DNA amount from 100 ng to 1 ug

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.).

Introduction

MagPure Stool DNA Kit is specially designed for high throughput DNA extraction from stool samples. It can get high purity microbial DNA from stool samples (≤200mg). This kit is based on magnetic beads purification and unique inhibiting factors adsorption technology, no phenol-chloroform extraction or alcohol precipitation. It can adsorb humic acid and other inhibiting factors in the solution efficiently. DNA can be directly used for downstream applications such as PCR, Viral DNA testing, bacterial DNA testing, etc.

Details

Specifications

Principle

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Lysozyme. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.

Advantages

• Fast – several samples can be extracted in 60 minutes (after digestion)
• High purity – unique adsorbent can completely remove inhibitory factors
• High recovery – DNA can be recovered at the level of PG

Kit Contents

Storage and Stability

MagPure Particles, RNase A and Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

MagPure Stool DNA Kit is specially designed for high throughput DNA extraction from stool samples. It can get high purity microbial DNA from stool samples (≤200mg). This kit is based on magnetic beads purification and unique inhibiting factors adsorption technology, no phenol-chloroform extraction or alcohol precipitation. It can adsorb humic acid and other inhibiting factors in the solution efficiently. DNA can be directly used for downstream applications such as PCR, Viral DNA testing, bacterial DNA testing, etc.

Overview

• Use with Norgen’s TaqMan Probe/Primer and Control Sets
• For any PCR application where the input template quality is important to avoid false negatives
• Convenience and time savings
• Cost efficient
• High sensitivity

TaqMan 2x PCR Master Mix – Cat. 28340

Norgen’s TaqMan 2x PCR Master Mix is a ready-to-use TaqMan 2x PCR Master Mix solution that contains a PCR internal control which can be detected by HEX/VIC channel in a real-time PCR machine. By detecting the internal control users can validate the DNA template quality, thereby preventing any false negatives in the PCR results. The user needs only to add template, target TaqMan primer/probe mix and water to set up the TaqMan real-time PCR. 

PCR Control: 
TaqMan 2x PCR Master Mix contains PCR control primers/probe (HEX/VIC) and PCR control template. The PCR control reaction in the TaqMan 2x PCR Master Mix is optimized to not interfere with target amplification. The fluorescence of the target probe should not be HEX/VIC.

Details

Reagents Supplied – Cat # 28341

• TaqMan 2x RT-PCR Master Mix (3 Vials, 100 Reactions) – Sufficient reagent for 100 x 20 µL reactions

Storage Conditions and Product Stability
Norgen’s TaqMan 2x PCR and 2x RT-PCR Master Mixes should be stored at -20ºC. For everyday use an aliquot can be stored at 4ºC for up to three months. The Master Mix is stable for multiple freeze-thaw cycles. When stored at the proper temperature this reagent is stable for at least 1 year.