Overview

• Convenient ready-to-use solution
• High sensitivity and yield
• Robust amplification
• Available in 3 convenient sizes: 100, 200 and 500 reactions (20 µL vol)
• This product is also a recommended add-on to be used with Norgen’s COVID-19 Primer and Probe Solutions and COVID-19 Positive Control 

Norgen’s 2X One-Step RT-PCR Master Mix is a ready-to-use solution that contains components required for RT-PCR amplification of RNA templates. The mix includes M-MuLV reverse transcriptase, Taq DNA polymerase, dNTPs, reaction buffer, MgCl₂, KCl, and a PCR enhancer/stabilizer. The user needs only to add the template, the primer set and water to the Master Mix to set up the RT-PCR reaction. This convenient 2X One-Step RT-PCR Master Mix reduces the time required to set up PCR reactions and reduces the possibility of contamination, particularly when preparing large numbers of reactions. The optimized master mix allows for robust amplification of RNA templates with high yields of PCR products.

Taq DNA Polymerase is a highly thermostable DNA polymerase that possesses a 5´→ 3´ polymerase activity and a very low 5´→ 3´ exonuclease activity. The source of Taq included with Norgen’s 2X One-Step RT-PCR Master Mix is an E. coli strain with a cloned Taq DNA Polymerase gene from Thermus aquaticus YT-1. M-MuLV Reverse Transcriptase is an RNA-directed DNA polymerase that can synthesize a complementary DNA strand initiating from a primer using either RNA (cDNA synthesis) or single-stranded DNA as a template. The source of the Reverse Transcriptase included with Norgen’s 2X One-Step RT-PCR Master Mix is an E. coli strain with a cloned reverse transcriptase gene from M-MuLV.

Norgen’s 2X One-Step RT-PCR Master Mix is available in 3 convenient sizes:

Cat # 28113 – 100 reactions (sufficient for 100 reactions x 20 µL reaction volume)
Cat # 28114 – 200 reactions (sufficient for 200 reactions x 20 µL reaction volume)
Cat # 28115 – 500 reactions (sufficient for 500 reactions x 20 µL reaction volume)

Details

Supporting Data

Figure 1 / 3

Previous

Next

Click for expanded view

Reagents supplied – Cat # 28113

• 2X One-Step RT-PCR Master Mix (1 Vials, 100 Reactions) – Sufficient reagent for 100 x 20 µL reactions

Storage Conditions and Product Stability
2X One-Step RT-PCR Master Mix should be stored at -20°C. For everyday use an aliquot can be stored at 4°C for up to 3 months. Repeated freeze-thaw cycles are not recommended. When stored at the proper temperature this reagent is stable for at least 1 year.

The NGS Library Quantification Standards with PCR Primers (for illumina platform) were developed for quantification of the NGS library concentration for illumina sequencing platform. It is comprised of Library Standards (six 10-fold dilutions) and a primer mix.

Quantification of the NGS library of the amplifiable molecules is critical for the quality of the sequencing data. Adequate library concentration will maximize sequencing capacity. Poor library concentration results in either low or high cluster density on the flow cell, which can lead to low sequencing capacity.

It is not accurate to measure the concentration of NGS library with standard DNA quantification methods such as spectrophotometer or fluorometer. QPCR is the best way for library quantification with high consistency and reproducibility of library quantitation.

BioDynami Library Quantification Standards with PCR Primers (for illumina platform): Amplification curve of 6 standards.

Our reagent is a highly sensitive, Real time PCR-based quantification that specifically designed for NGS libraries using illumina sequencing platform. The amplification uses illumina adaptor sequences as primers, and only the fully adaptor-ligated libraries will be amplified.  Therefore the reagent provides an accurate estimation of the library concentration based on true sequenceable illumina libraries. In addition, this kit can also be used for confirmation of ligation reaction after completion of library preparation.

Our library quantification standards is compatible with commercial SYBR Green based QPCR reagents. This makes it more flexible for scientists who want to use their real time PCR reagent. Quantification of library concentration is achieved by comparison with a standard curve generated from the Library Standards.

The NGS Library Quantification Standards with PCR Primers (for illumina platform) were developed for quantification of the NGS library concentration for illumina sequencing platform. It is comprised of Library Standards (six 10-fold dilutions) and a primer mix.

Introduction

The kit offers the unique feature to isolate total RNA including small RNA and DNA from serum and plasma without the need to resort to the cumbersome phenol/chloroform extraction or a time consuming proteinase digest. RNA purified using the kit is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA and miRNA from 0.2-0.6ml serum and plasma
Applications
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type
Sample amount
Yield
Elution volume
Time per run
Liquid carrying volume per column
Binding yield of column

Principle

HiPure RNA technology simplifies total RNA isolation. The sample material is denatured in Lysis Buffer. The protein is then precipitated by Protein Precipitation Solution and pelleted by centrifugation. After the removal of protein the binding conditions for nucleic acids are adjusted by adding isopropanol. Total nucleic acids are bound to the column. Optionally, DNA can be removed by an on-column DNase digest. The remaining nucleic acids are washed and eluted with minimal amounts of RNase-Free water.

Kit Contents

Contents	R431402	R431403
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	100	2 x 250
2ml Collection Tubes	100	2 x 250
Buffer CFL	6 ml	30 ml
Buffer CFP	1.8 ml	10 ml
Buffer RWC	20 ml	80 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	60 ml

Storage and Stability

The kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

The kit offers the unique feature to isolate total RNA including small RNA and DNA from serum and plasma without the need to resort to the cumbersome phenol/chloroform extraction or a time consuming proteinase digest. RNA purified using the kit is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.