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

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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.

Introduction

The Kit provides fast purification of high-quality RNA from plants, cell, tissues, and yeast using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol/chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or precipitation with isopropanol or LiCl. RNA purified using the RaPure Total RNA Purification System can be used 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 from <150 mg simple plant sample without chloroform
Applications	RT-PCR, qPCR, Northern hybridization, second generation sequencing, nucleic acid protection, in vitro translation
Purification method	Mini spin column
Purification technology	Silica technology, DNA filtration technology
Process method	Manual (centrifugation or vacuum)
Sample type	Economic crops
Sample amount	≤150 mg
Elution volume	≥30μl
Time per run	≤25 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

The Kit isolates total RNA from up to 150mg plant tissue. A short workflow enables RNA isolation with genomic DNA removal in less than 25 min. Samples are first lysed and homogenized. The lysate is passed through a DNA Mini column, ethanolis added to the flow-through, and the sample is applied to an RNA column. RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in as little as 30µl water using the Kit.

Advantages

• Efficient removal of DNA – unique genomic DNA removal column without DNase treatment
• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Fast – several samples can be extracted in 25 minutes by column method
• Safe – no phenol chloroform extraction required
• Sensitive – RNA can be recovered at the level of PG
• Broad spectrum – various types of plant samples can be processed by diversity of operating procedures

Kit Contents

Contents	R415102	D415103
Purification Times	50 Preps	250 Preps
HiPure DNA Mini Column II	50	250
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	100	2 x 250
Buffer RLC	50 ml	200 ml
Buffer PRC1	50 ml	200 ml
Buffer RW1	50 ml	200 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

The Kit can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. Make sure that all buffers are at room temperature when used. During shipment, crystals or precipitation may form inthe Buffer RLC/PRC1. Dissolve by warming buffer to 37°C.

Purchase Guide

1. When dealing with woody or uncommon samples, R4150 is recommended first. R4150 contains two polysaccharide/polyphenol lysis buffer, which is the most universal product.

2. R4151 is recommended for handling common economic crop samples for the first time. Strong lysis solution can be used to process easy-extraction samples. The amount of corn or rice leaves samples can reach up to 300mg.

3. R4165 adopts CTAB/chloroform method, which can also handle a large number of difficult-to-extraction plants, but requires contact with chloroform substitutes, which is less safe than other kits. This kit uses DNase Ⅰ to remove DNA, which is also a good choice for extracting polysaccharide/polyphenol-rich plant samples.

4. R4014 is recommended for fruit/starch plant samples, which uses improved trizol pre-treatment, single column operation and is more economical.

Select the right purification kit to get impactful results：

Ⅰ. Purchase guide of Magen kits based on sample

Ⅱ. Cross purchase guide for Magen kits vs Other brands

For any customized product or OEM service, please contact us.

The Kit provides fast purification of high-quality RNA from plants, cell, tissues, and yeast using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol/chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or precipitation with isopropanol or LiCl. RNA purified using the RaPure Total RNA Purification System can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.

Genomic DNA Extraction Kit (HMW, Magnetic Beads)

The Genomic DNA Extraction Kit (HMW, Magnetic Beads) provides a reliable and fast process for extracting high molecular weight (HMW) genomic DNA from cells, blood, and tissues using Solid Phase Reversible Immobilization (SPRI) magnetic beads. With our proprietary magnetic beads technology, the kit eliminates the tedious centrifuge steps for columns. The kit provides a reliable and simple approach for high-quality genomic DNA isolation with fast magnetic response time and high binding capacity.

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Cat.# 50014 Genomic DNA Extraction Kit for Cells (HMW, Magnetic Beads)
Cat.# 50015 Genomic DNA Extraction Kit for Blood (HMW, Magnetic Beads)
Cat.# 50016 Genomic DNA Extraction Kit for Tissues (HMW, Magnetic Beads)

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The extracted HMW genomic DNA size ranges are dependent on the beads resuspension: 50-150 kb by tube tapping and 40-100 kb by tube vortexing. Purified DNA is recovered at high yield and high purity without RNA contamination. The typical purity ratios of A260/A280 are around 1.8-2.0, and A260/A230 are around 2.2-2.5. Purified HMW genomic DNA is suitable for applications such as long-read sequencing, linked-read genome assembly, long range PCR, optical mapping, and other general applications.

Features

• High molecular weight DNA: 50 kb to 150 kb
• High purity
• Simple magnetic beads method
  • No centrifuge needed
  • No column needed
  • No vacuum needed

A portion of the extracted genomic DNA samples were loaded on a PFGE gel with a DNA ladder indicated. Sample A: liver tissue; Sample B: intestine tissue; Sample C: whole blood; Sample D: cultured 293T cells.

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Cultured Cell samples

Cultured cells are collected and are resuspended in a buffer and then lysed with a lysis buffer, then mixed with beads to bind genomic DNA. The samples are mixed with a buffer and after washing steps, genomic DNA is eluted in the Elution Buffer. The isolated genomic DNA with the magnetic beads is free of contamination such as RNA, proteins, salts, and other impurities.

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Blood samples

Whole blood is resuspended in the RBC buffer to remove RBC. The remaining leucocytes are lysed with a lysis buffer, then mixed with beads to bind genomic DNA. The samples are mixed with a buffer and after washing steps, genomic DNA is eluted in Elution Buffer. The isolated genomic DNA with the magnetic beads is free of contamination such as RNA, proteins, salts, and other impurities.

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Tissue samples

Tissues are homogenized and lysed, then mixed with beads to bind genomic DNA. The samples are mixed with a buffer and after washing steps, genomic DNA is eluted in Elution Buffer. The isolated genomic DNA with the magnetic beads is free of contamination such as RNA, proteins, salts, and other impurities.