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

Magnetic Beads (Short Oligo Purification)

Solid Phase Reversible Immobilization (SPRI) beads are a simple and effective reagent for DNA purification, but not for short oligo purification. The beads are paramagnetic particles coated with carboxyl groups that can reversibly bind to nucleic acid. However, SPRI beads can only purify DNA/RNA fragments that are 100 base pairs or longer. DNA/RNA fragments shorter than 100 base pairs are not effectively recovered.

Oligo purification can also be performed using spin column-based technology. The oligo size limitation for recovery is around 20 nt, as oligos under 20 nt have a very low recovery rate.

We have developed Magnetic Beads (Short Oligo Purification) Kit for short oligo purification. Our proprietary bead technology enables the recovery of oligos as short as 6 nt. 80% of the 6 nt oligos and 90% of the > 8 nt oligos can be recovered. The reagent also effectively removes impurities and unwanted components such as salts, proteins, dNTPs, detergents, and other contaminants. The magnetic bead reagents are RNase free, and can be used for both DNA and RNA applications.

Recovery rates of of short oligos. Oligos with different sizes were used as input. BioDynami Short Oligo Quantification Kit (Cat. # 40046) was used to quantify the recovery rates with modifications.

Features

• Effective purification of short oligos
  • • 6 nt oligos: 80% recovery rate
    • >8 nt oligos: >90% recovery rate
• Removal of impurities and unwanted reaction components

Solid Phase Reversible Immobilization (SPRI) beads are a simple and effective reagent for DNA purification, but not for short oligo purification. The beads are paramagnetic particles coated with carboxyl groups that can reversibly bind to nucleic acid. However, SPRI beads can only purify DNA/RNA fragments that are 100 base pairs or longer. DNA/RNA fragments shorter than 100 base pairs are not effectively recovered.

K-MBG4

SKU: 700004319

100 assays (manual) / 400 assays (auto-analyser)

Content:	(Malt β-glucanase) 100 assays (manual) / 400 (auto-analyser) Or (Lichenase) 100 / 200 assays (manual) / 330 (auto-analyser)
Shipping Temperature:	Ambient
Storage Temperature:	Short term stability: 2-8oC, Long term stability: See individual component labels
Stability:	> 2 years under recommended storage conditions

Analyte:	β-Glucanase/Lichenase
Assay Format:	Spectrophotometer, Auto-analyser
Detection Method:	Absorbance
Wavelength (nm):	400
Signal Response:	Increase
Limit of Detection:	(Malt β-glucanase) 4.3 x 10-4 U/mL Or (Lichenase) 9.1 x 10-5 U/mL
Reproducibility (%):	~ 3%
Total Assay Time:	(Malt β-glucanase) ~ 20 min Or (Lichenase) ~ 10 min
Application examples:	Crude malt extracts, industrial enzyme preparations.
Method recognition:	Novel method

The MBG4 reagent contains a single substrate, namely 4,6-O-benzylidene-2-chloro-4-nitrophenyl-β-(3¹-β-D-cellotriosyl-glucoside) (BCNPBG4). The benzylidene acetal group prevents any hydrolytic action by exo-acting hydrolytic enzymes such as β-glucosidase or cellobiohydrolase.

Mixed linkage β-glucanase (endo-1,3:1,4-β-glucanase) / lichenase (EC 3.2.1.73) acts specifically to release 2-chloro-4-nitrophenol (CNP) from this substrate. The rate of release of CNP is directly related to the β-glucanase/lichenase activity in a sample. The reaction is terminated and the phenolate colour is developed on addition of Tris buffer solution (pH = 10.0).

Note that the substrate is not hydrolysed by β-glucosidase or cellobiohydrolase. The substrate can be hydrolysed by certain endo-cellulases (e.g. Trichoderma sp.) but this does not result in an increase in absorbance.

Discover more assay kits for enzyme activity measurement.

Data calculators are located in the Documents tab.

Advantages

• Very cost effective 
• All reagents stable for > 2 years 
• Specific for endo-1,3:1,4-β-glucanase/lichenase 
• Simple, convenient, rapid assay 
• Well suited to automation 
• Malt flour standard and lichenase standard included

The MBG4 reagent contains a single substrate, namely 4,6-O-benzylidene-2-chloro-4-nitrophenyl-β-(31-β-D-cellotriosyl-glucoside) (BCNPBG4). The benzylidene acetal group prevents any hydrolytic action by exo-acting hydrolytic enzymes such as β-glucosidase or cellobiohydrolase.