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 # 28114

• 2X One-Step RT-PCR Master Mix (2 Vials, 100 Reactions each) – Sufficient reagent for 200 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.

Introduction

This product provides fast and easy methods for purification of total DNA for reliable PCR and southern blotting. Total DNA can be purified from bacterial culture, body fluids, food and fermentation.

Details

Specifications

Principle

This product is based on silica Column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid is adsorbed on the membrane, while proteinis not adsorbed and is removed with filtration. After washing proteins and other impurities, Nucleic acid was finally eluted with low-salt buffer (10mmTris, pH9.0, 0.5mm EDTA). This product carries 0.1-0.2mm acid glass beads, which can improve the wall-breaking effect of lysozyme resistant bacteria, the success rate and DNA yield through physical bead grinding.

Advantages

• Fast – several samples can be extracted in 40 minutes (after digestion)
• High purity – purified DNA can be directly used in various downstream applications
• Good repeatability – silica technology can obtain ideal results every time
• High recovery – DNA can be recovered at the level of PG
• Broad spectrum – it can deal with a variety of bacteria, including Gram-positive bacteria that are difficult to disrupt
• Sufficient components – this kit has carried lysozyme, protease K, RNase A and glass beads

Kit Contents

Storage and Stability

Lysozyme and Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 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.

This product provides fast and easy methods for purification of total DNA for reliable PCR and southern blotting. Total DNA can be purified from bacterial culture, body fluids, food and fermentation.