N-(Propargyl-PEG2)-N-bis(PEG3-t-butyl ester)

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 # 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.

Description

The oasig freeze drying process stabilises all of the active components allowing these reagents to be shipped and stored at room temperature. This hugely simplifies the logistics of purchasing, shipping and using the technology. Whether you are in a sophisticated laboratory in Texas or a mobile field hospital in Timbuktu we can supply complete qPCR kit and reagent packages to your door quickly and cheaply via standard shipping methods without the need for dry ice or a cold chain of any sort.

The performance of the reagents is second to none. We are confident that you will find excellent data quality and even see an improvement in data quality versus many traditional frozen master mixes.

Primerdesign real-time PCR reagents are manufactured to the highest standards within our ISO9001:2008 and ISO13485:2012 certified quality management laboratory environment.

150 reaction pack of freeze dried oasig Lyophilised OneStep RT-qPCR Mastermix. 3 x 50 reaction vials and resuspension buffer.

Introduction

Intended Use

For the isolation and differentiation of sorbitol-negative Escherichia coli serotype O157 from food and animal feed and other materials

Principle and Interpretation

Enzymatic digest of casein and enzymatic digest of animal tissue provides nitrogen source, vitamins and growth factors; sodium chloride maintains balanced osmoticpressure; sorbitol is a fermentable sugar; No. 3 bile salt and crystal violet inhibit Gram-positive bacteria, potassiumtellurite inhibits non-o157 Escherichia coli, and cefixime inhibits Proteus; neutral red is a pH indicator; and agar is the coagulant of the culture medium.

Formulation

Ingredients	/liter
Enzymatic digest of casein	17.0g
Enzymatic digest of animal tissue	3.0g
Sorbitol	10.0g
Bile salt No.3	1.5g
Sodium chloride	5.0g
Neutral red	0.03g
Crystal violet	0.001g
Agar	15.0g
pH 7.1±0.2 at 25°C

Preparation

Weigh 51.5g of this product, add 1L of distilled water or deionized water, stir, heat and boil until completely dissolved, dispense into Erlenmeyer flasks, and sterilize at 121℃ for 15min. Melt the sterilized culture medium and cool it to about 45℃, add 1 tube of supporting reagent (SR0240) (0.25mg potassium tellurite and 0.005mg Cefixime) to every 100mL of basal culture medium, mix and pour into the plate.

Quality Control

Cultural characteristics observed after incubation at 35-37°C for 18-24hours

Quality control strains	Growth	Colony color
Escherichia coli ATCC25922	PR≥0.7	pale pink to red
Escherichia coli 0157:H7 NCTC12900	PR≥0.7	colorless
Staphylococcus aureus ATCC 25923	Total inhibition	–

Sorage and Shelf Life

Keep container tightly closed, store in a cool, dry place, away from bright light. Storage period of 3 years.

Precautions

1. When weighing the dehydrated medium, please wear masks to avoid causing respiratory system discomfort

2. Keep container tightly closed after using to prevent clumping.

Waste Disposal

Microbiological contamination was disposed by autoclaving at 121°C for 30 minutes.

Intended Use For the isolation and differentiation of sorbitol-negative Escherichia coli serotype O157 from food and animal feed and other materials Principle and Interpretation Enzymatic ……