4x12x7ml Swing Buckets Rotor

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.

Overview

• Sequentially purify total RNA and total proteins from a single sample
• Kit includes a gDNA elimination column  
• No sample splitting required
• No phenol step required for efficient isolation
• Ideal for small or difficult to obtain samples
• Purify RNA and proteins from cultured animal cells, tissues, blood, bacteria, yeast, fungi or plants
• Rapid and efficient spin column procedure
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

This kit provides a rapid, single column method for the isolation and purification of total RNA (including miRNA) and proteins sequentially from a single sample of cultured animal cells, tissues, blood, bacteria, yeast, fungi or plants. The total RNA and proteins are both column purified in under 25 minutes using a single column.

Purified RNA is of a high quality and yield, and is suitable for NGS, RT-qPCR and microarrays.  

This kit eliminates DNA efficiently using a gDNA removal column.

Proteins are eluted in buffer and are ready for downstream applications such as Western Blots, Mass Spec and ELISA.  The proteins will not require precipitation, resuspending of pellets, or any further cleaning.

This kit is ideal for researchers who are interested in studying the transcriptome and proteome of a single sample, such as for studies of microRNA profiling, gene expression including gene silencing experiments or mRNA knockdowns, studies involving biomarker discovery, and for characterization of cultured cell lines. Norgen’s RNA/Protein Purification Plus Kit is especially useful for researchers who are isolating macromolecules from precious, difficult to obtain or small samples such as biopsy materials or single foci from cell cultures, as it eliminates the need to fractionate the sample. Furthermore, analysis will be more reliable since the RNA and proteins are derived from the same sample, thereby eliminating inconsistent results. The purified macromolecules are of the highest purity and can be used in a number of different downstream applications.

Protocol

Details

Supporting Data

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Kit Specifications
Maximum Column Binding Capacity	50 μg for RNA
Maximum Column Binding Capacity	200 μg for Protein
Maximum Column Loading Volume	650 μL
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Time to Complete 10 Purifications	30 minutes
Average Yields*: HEK 293 Cells (1 x 106 cells) HEK 293 Cells (1 x 106 cells) Liver (15 mg) Liver (15 mg)	10-15 μg RNA 70-100 μg protein 30-35 μg RNA 100-150 μg protein

* Average yields will vary depending upon a number of factors including species, growth conditions used and developmental stage.

Storage Conditions and Product Stability
The Protein Loading Dye should be stored at -20°C after the addition of DL-Dithiothreitol (DTT). All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.

Component	Cat. 48200 (50 preps)
Buffer SKP	40 mL
Wash Solution A	38 mL
Elution Solution A	6 mL
Wash Solution C	30 mL
Binding Buffer A	8 mL
Elution Buffer C	4 mL
Protein Neutralizer	4 mL
Protein Loading Dye	2 mL
gDNA Removal Columns	50
RNA/Protein Purification Columns	50
Collection Tubes	150
Elution Tubes (1.7 mL)	100
Product Insert	1