Overview

• Convenient
• One cDNA Synthesis, Multiple microRNAs and microRNA-targets analyzed
• Time Savings
• Cost Efficient
• High Sensitivity and Yield
• Robust Enzyme
• Available in 12 or 50 reaction size

Norgen’s microScript microRNA cDNA Synthesis Kit is an all-in-one, ready-to-use product for the reverse transcription of microRNA from either Total RNA preparations or enriched microRNA preparations. The kit contains the 2x Reaction Mix and the microScript microRNA Enzyme Mix. The kit utilizes Norgen’s microScript Reverse Transcriptase, a mutant version of Moloney Murine Leukemia Virus (M-MuLV) Reverse Transcriptase. It has reduced RNase H activity and increased thermal stability.

The workflow of Norgen’s microScript microRNA cDNA Synthesis Kit involves a simple, single-tube set-up by the mixing of 2x Reaction Mix, Enzyme Mix and the RNA template. The reaction can then be carried out in a thermocycler. A poly (A) tail is first added to the RNA template, followed by cDNA synthesis using an adapter primer. In addition to the ease-of-use, the single-tube set-up provides superb consistency and sensitivity. The cDNA could be used in a PCR or qPCR amplification using a Universal PCR Reverse Primer and the forward primer that contains the sequence of the microRNA of interest. A single cDNA preparation could be used for PCR amplification of a number of different microRNAs. In addition, the cDNA preparation could be used for PCR or qPCR detection (using gene-specific forward and reverse primers) of mRNA or large RNA if total RNA preparation was the starting template. This could allow for parallel evaluation of expression level of microRNAs and microRNA-targets.

Details

Supporting Data

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Storage Conditions and Product Stability
Norgen’s microScript microRNA cDNA Synthesis Kit components should be stored at -20°C. These reagents should remain stable for at least 1 year in their unopened containers.

Component	Cat. 54415 (12 rxns)	Cat. 54410 (50 rxns)
microScript microRNA Enzyme Mix	12 μL	50 μL
2x Reaction Mix	120 μL	500 μL
Universal PCR Reverse Primer	60 μL	250 μL
Nuclease-Free Water	1.25 mL	2 x 1.25 mL
Product Insert	1	1

All products are offered in bulk quantities and can be pre-dispensed in PCR-strips, PCR-plates, cartridges etc… Significant discounts are available.

Customized products also including primers & probes can be provided on demand as well. Please contact us to receive an individual quotation. Get in touch! 

All products are offered in bulk quantities and can be pre-dispensed in PCR-strips, PCR-plates, cartridges etc… Significant discounts are available.

Customized products also including primers & probes can be provided on demand as well. Please contact us to receive an individual quotation. Get in touch!

Introduction

This product is suitable for rapid extraction of total DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from tissue / blood / body fluid / swab /dry blood spots
Applications	PCR, qPCR, southern bolt and virus detection, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Tissue, cell, blood, saliva, swab, blood spot, semen and other clinical samples
Sample amount	Solid tissue : 1-10mg, Anticoagulant blood : 200µl
Yield	1 – 15µg
Elution volume	≥20μl
Time per run	30 – 60 minutes
Liquid carrying volume per column	750µl
Binding yield of column	100µg

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 protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).

Advantages

• High quality DNA – meet a variety of downstream applications, including PCR, qPCR, enzyme digestion, hybridization, etc.
• Fast – without separation of leukocytes, organic extraction or ethanol precipitation
• Simple – all nucleic acids can be obtained by direct digestion
• Wide applicability – It can handle various liquid samples, animal tissues and cultured cells

Kit Contents

Contents	IVD3018
Purification Times	100
HiPure DNA Mini Columns I	100
2ml Collection Tubes	2 x 100
Buffer ATL	60 ml
Buffer AL	60 ml
Buffer GW1	44 ml
Buffer GW2	50 ml
Proteinase K	60 mg
Protease Dissolve Buffer	5 ml
Buffer AE	15 ml

Storage and Stability

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 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.

Experiment Data

This product is suitable for rapid extraction of total DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.