Overview

• Convenient – With the ready-to-use Master Mix, the user needs only to add template to the master mix and enzyme in order to set up the reverse transcriptase reaction
• Time Savings – Set up RT reactions in a shorter time since less pipetting steps are required
• Cost Efficient – No need to buy separate enzymes, dNTPs and buffers. All are included with the ready-to-use Master Mix kits
• High Sensitivity and Yield – the optimized Master Mix allows for highly sensitive amplifications with high yields of PCR products
• Robust Enzyme – broad range of working temperatures from 37°C to 60°C. Capable of amplifying difficult templates with a high degree of reproducibility

TruScript Reverse Transcriptase is Available in Three Convenient Formats:

1. TruScript Reverse Transcriptase Kit (Cat.#54440)

This kit contains 5X RT Buffer and a vial of TruScript Enzyme Mix (200 units/µL). This enzyme can be used for reverse-transcription reactions with any user-supplied primers.

2. TruScript First Strand cDNA Synthesis Kit (Cat.#54420)

This is an all-in-one, ready-to-use product for simple set-up of reverse transcription of total RNA (both poly A- or non-poly A-containing transcripts).

The kit contains the 2x Reaction Mix and the TruScript Enzyme Mix. The 2x Reaction Mix contains a blend of buffer, nucleotides and primers (oligo dT and random hexamers) for effective cDNA synthesis from total RNA transcripts.

3. TruScript First Strand cDNA Synthesis Kit for mRNA (Cat.#54400)

This is an all-in-one, ready-to-use product for simple set-up of reverse transcription of messenger RNA (poly A-containing transcripts).

The kit contains the 2x Reaction Mix and the TruScript Enzyme Mix. The 2x Reaction Mix contains a blend of buffer, nucleotides and oligo dT primer for the effective cDNA synthesis from total RNA transcripts or enriched mRNA sample.

Which TruScript Kit is Best for Your Needs?

	TruScript Reverse Transcriptase	TruScript First Strand cDNA Synthesis Kit for mRNA	TruScript First Strand cDNA Synthesis Kit
Kit contains only reverse transcriptase enzyme and buffer (no primers or other buffer components)
Your template is enriched mRNA
Your template is total RNA (poly A OR non-poly A-containing transcripts)
Kit contains oligo dT
Kit contains both oligo-dT primer and Random Hexamers
Kit contains reaction buffer with nucleotides and primer		with oligo-dT	with oligo-dT and random hexamers
You have your own first strand synthesis primer
Your template is microRNA (using your own primers)
	Cat.#54440	Cat.#54400	Cat.#54420

Details

Supporting Data

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TruScript First Strand cDNA Synthesis Kit (Cat.# 54420) – Kit Components

1. TruScript Enzyme Mix
2. 2x Reaction Mix
3. Nuclease-Free Water 
4. Product Insert

Storage Conditions and Product Stability

Norgen’s TruScript Reverse Transcriptase and the 5x RT Buffer should be stored at -20°C.  These reagents should remain stable for at least 1 year in their unopened containers.

Component	Cat. 54440 (10,000 units)	Cat. 54420 (50 rxns)	Cat. 54410 (50 rxns)
TruScript™ Reverse Transcriptase (200 units/ μL)	50 μL	–	–
5x RT Buffer	1 mL	–	–
TruScript™ Enzyme Mix	–	100 μL	100 μL
2x Reaction Mix	–	500 μL	–
2x Reaction Mix for mRNA	–	–	500 μL
Nuclease-Free Water	1.25 mL	1.25 mL	1.25 mL
Product Insert	1	1	1

Introduction

With the development of molecular biology, stool, a new non-invasive sample, has been widely used in the research of animal molecular genetics, population ecology, behavioral ecology and some intestinal disease diagnosis. Stool samples includes gut microbial DNA, food residue sample DNA, and alimentary tract exfoliated cell DNA.

The primary problem encountered when using stool sample for molecular biology research is the low content of exfoliated cells in the digestive tract and a certain degree of degradation of genetic material in stool. Another issue in molecular scatology research based on PCR is the presence of a large number of inhibitors in stool that can affect Taq enzyme activity, leading to downstream detection inactivation. These inhibitors include polysaccharides, plant polysaccharides, bile acids, bile salts, bile pigments, digestive juices, mucus, etc. Therefore, selecting appropriate extraction methods to obtain high-quality DNA is the key to successful downstream detection of stool DNA.

At present, the pretreatment methods used in the laboratory, such as phenol/chloroform extraction, cetyltrimethyl bromide (CTAB) lysis, and guanidine isothiocyanate lysis, lack universality in different species, and the success rate of extracting DNA for PCR amplification is also very low. The HiPure Stool DNA Kit provided by Magen Company has opened up a new approach for DNA extraction from stool samples with good universality, high cost-effectiveness, high yield and purification. The reagent kit adopts a unique solution system and inhibitory factor adsorbent, which can efficiently remove various impurities in stool samples. The purified DNA can be directly used for PCR, quantitative PCR and other applications.

This product allows rapid and reliable isolation of high-quality genomic DNA from various stool samples. Up to 100 mg soil samples can be processed in 60 minute. The system combines the reversible nucleic acid binding properties of HiPure matrix with the speed and versatility of spin column technology to eliminate PCR inhibiting compounds such as humic acid from soil samples. Purified DNA is suitable for PCR, restriction digestion, and next-generation sequencing. There are no organic extractions thus reducing plastic waste and hands-on time to allow multiple samples to be processed in parallel.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from 50-100mg stool samples
Applications	PCR, Southern Blot, enzyme digestion and NGS, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Stool
Sample amount	50-100mg
Yield	3-15μg
Elution volume	≥30μl
Time per run	≤60 minutes
Liquid carrying volume per column	750μl
Binding yield of column	100μg

Principle

Stool sample is homogenized and then treated in a specially formulated buffer containing detergent to lyse bacteria, yeast, and fungal samples. Humic acid, proteins, polysaccharides, and other contaminants are removed using our proprietary Absorber Solution. Binding conditions are then adjusted and the sample is applied to a DNA Mini Column. Two rapid wash steps remove trace contaminants and pure DNA is eluted in low ionic strength buffer. Purified DNA can be directly used in downstream applications without the need for further purification.

Advantages

• High purity – unique adsorbent can completely remove inhibitory factors
• High concentration – maximum extraction of total DNA from stool samples
• High recovery – DNA can be recovered at the level of PG
• Good repeatability – silica technology can obtain ideal results every time

Kit Contents

Contents	D314102	D314103
Purification Times	50 Preps	250 Preps
HiPure DNA Mini Columns II	50	250
2ml Collection Tubes	50	250
2ml Bead Tubes	50	250
Proteinase K	24 mg	120 mg
Protease Dissolve Buffer	1.8 ml	10 ml
Buffer SPL	40 ml	200 ml
Buffer PCI	40 ml	200 ml
Buffer AL	20 ml	80 ml
Buffer GW1	22 ml	88 ml
Buffer GW2	20 ml	2 x 50  ml
Buffer AE	15 ml	30 ml

Storage and Stability

Proteinase K and Buffer PCI 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 at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2–8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

With the development of molecular biology, stool, a new non-invasive sample, has been widely used in the research of animal molecular genetics, population ecology, behavioral ecology and some intestinal disease diagnosis. Stool samples includes gut microbial DNA, food residue sample DNA, and alimentary tract exfoliated cell DNA.

The primary problem encountered when using stool sample for molecular biology research is the low content of exfoliated cells in the digestive tract and a certain degree of degradation of genetic material in stool. Another issue in molecular scatology research based on PCR is the presence of a large number of inhibitors in stool that can affect Taq enzyme activity, leading to downstream detection inactivation. These inhibitors include polysaccharides, plant polysaccharides, bile acids, bile salts, bile pigments, digestive juices, mucus, etc. Therefore, selecting appropriate extraction methods to obtain high-quality DNA is the key to successful downstream detection of stool DNA.

At present, the pretreatment methods used in the laboratory, such as phenol/chloroform extraction, cetyltrimethyl bromide (CTAB) lysis, and guanidine isothiocyanate lysis, lack universality in different species, and the success rate of extracting DNA for PCR amplification is also very low. The HiPure Stool DNA Kit provided by Magen Company has opened up a new approach for DNA extraction from stool samples with good universality, high cost-effectiveness, high yield and purification. The reagent kit adopts a unique solution system and inhibitory factor adsorbent, which can efficiently remove various impurities in stool samples. The purified DNA can be directly used for PCR, quantitative PCR and other applications.

This product allows rapid and reliable isolation of high-quality genomic DNA from various stool samples. Up to 100 mg soil samples can be processed in 60 minute. The system combines the reversible nucleic acid binding properties of HiPure matrix with the speed and versatility of spin column technology to eliminate PCR inhibiting compounds such as humic acid from soil samples. Purified DNA is suitable for PCR, restriction digestion, and next-generation sequencing. There are no organic extractions thus reducing plastic waste and hands-on time to allow multiple samples to be processed in parallel.

Description

Specifications

Clone	IHC026
Source	Rabbit Monoclonal
Positive Control	Colon Cancer
Dilution Range	1:200

MutS Homolog 6 (MSH6) is a protein involved in the mismatch repair pathway. This protein is commonly associated with hereditary non-polyposis colorectal cancer, and mutations in this gene are correlated with the development of sporadic colorectal carcinoma. Studies have shown that mutations in MSH6, when co-indicated with mutations in MSH1 and MSH2, contribute to the development of sporadic colorectal carcinoma. Use of Anti-MSH2 is optimized when paired with MSH6, MLH1, and PMS2 in an IHC panel.