Introduction

This product is specially designed for stool RNA extraction. The kit is suitable for extracting high-purity microbial or host cell RNA from ≤ 0.15g stool samples. The purified RNA can be directly used in RT-PCR and Northern hybridization.

Details

Principle

The Kit combines the speed and efficiency of silica-based technology with the convenient handling of magnetic particles for purification of total RNA. Samples are lysed and RNA is purified from lysates in one step through its binding to the silica surface of the particles in the presence of a chaotropic salt. The particles are separated from the lysates using a magnet and DNA is removed by treatment with RNase-free DNase. The magnetic particles are efficiently washed, and RNA is eluted in RNase-free water.

Kit Contents

Contents	R662601	R662602	R662603
Purification Times	48 Preps	96 Preps	5 x 96 Preps
MagPure RNA Particles	1.7 ml	4.0 ml	18 ml
DNase I	600 μl	2 x 600 μl	10 x 600 μl
DNase Buffer	30 ml	40 ml	200 ml
Buffer SPL	30 ml	60 ml	270 ml
Buffer PHC	30 ml	60 ml	270 ml
Buffer MCB*	9 ml	15 ml	75 ml
Buffer ALB3*	10 ml	20 ml	100 ml
Buffer GW1*	44 ml	66 ml	2 x 220 ml
Buffer RW2*	20 ml	50 ml	2 x 100 ml
RNase Free Water	10 ml	30 ml	120 ml
2ml Bead Tubes	48	96	5 x 96

Storage and Stability

MagPure RNA Particles should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, MagPure RNA Particles up to 8 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.

This product is specially designed for stool RNA extraction. The kit is suitable for extracting high-purity microbial or host cell RNA from ≤ 0.15g stool samples. The purified RNA can be directly used in RT-PCR and Northern hybridization.

Description 

ExcelRT™ Reverse Transcription Kit is a complete, efficient and convenient kit to synthesize high quality first strand cDNA. This kit contains ExcelRT™ Reverse Transcriptase, which is able to synthesize the first strand cDNA at 37~50°C. The ExcelRT™ Reverse Transcriptase is a recombinant Moloney Murine Leukemia Virus (M-MLV) reverse transcriptase, which is designed to reduce RNase H activity and create better thermal stability. This kit also contains RNAok™ RNase Inhibitor, which is active against RNase A, RNase B, and RNase C. This product is supplied with oligo (dT)₂₀ and random hexamers, which are used to synthesize cDNA from poly(A) tailed mRNA and total RNA, respectively. 

Features

• Contains all components for reverse transcription 
• High yield 
• Thermostable, up to 50°C, during first strand synthesis 
• High processivity, generating cDNA up to 8 kb 
• Reduced RNase H ribonuclease activity 

Application

• Generation of first strand cDNA from total RNA or mRNA.
• Suitable for generating cDNA from RNA with strong secondary structure which can be reduced at temperature up to 50°C.

Storage

-20°C for 24 months 

High yield

High sensitivity

Thermostable, up to 50°C, during first strand synthesis

 

Contents

ComponentVolume    ExcelRT™ Reverse Transcriptase (200 U/μl) 100 μl      RNase Inhibitor (20 U/μl)100 μl      5X RT Buffer (DTT)500 μl      dNTPs Mix (10 mM each)200 μl      Oligo (dT)20 (50 μM)100 μl      Random Hexamers (100 μM)100 μl      DEPC-Treated H2O  1 ml x 2

Storage Buffer

Reverse Transcriptase: 20 mM Tris-HCl (pH 7.5), 200 mM NaCl, 0.1 mM EDTA, 1 mM DTT, stabilizer and 50% (v/v) glycerol

RNase Inhibitor: 40 mM HEPES-KOH (pH 7.5), 100 mM KCl, 8 mM DTT, 0.1 mM EDTA, stabilizer and 50% (v/v) glycerol

5X RT buffer (DTT)

250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl₂ and 50 mM DTT

Storage

-20°C for 24 months 

ExcelRT™ Reverse Transcription Kit is a complete, efficient and convenient kit to synthesize high quality first strand cDNA. This kit contains ExcelRT™ Reverse Transcriptase, which is able to synthesize the first strand cDNA at 37~50°C. The ExcelRT™ Reverse Transcriptase is a recombinant Moloney Murine Leukemia Virus (M-MLV) reverse transcriptase, which is designed to reduce RNase H activity and create better thermal stability. This kit also contains RNAok™ RNase Inhibitor, which is active against RNase A, RNase B, and RNase C. This product is supplied with oligo (dT)20 and random hexamers, which are used to synthesize cDNA from poly(A) tailed mRNA and total RNA, respectively.