Shigella species

Overview

• Excellent separation and purification of cytoplasmic and nuclear RNA
• Convenient and fast spin column format
• High quality and yield of RNA
• Isolate full diversity of RNA (including microRNA) without phenol
• Purified RNA is ready for any application including RT-PCR, qRT-PCR, RNA-Seq, arrays and more
• Cytoplasmic RNA is free of DNA and ready for direct use in RT-PCR/qRT-PCR
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

This kit provides a rapid method for the isolation and purification of both cytoplasmic and nuclear RNA from cultured animal cells and small tissue samples. The kit can be used to isolate all sizes of RNA from the cytoplasmic and nuclear RNA fractions, including all small RNA species without any requirement for phenol.   Included in the kit are sufficient reagents to perform either 50 cytoplasmic RNA preparations or 25 cytoplasmic and 25 nuclear RNA preparations.   Ten samples can be processed in approximately 45 minutes.  This kit is also available in a 100 prep size.    

Background

In certain circumstances it is desirable to be able to isolate fractionated RNA as opposed to total RNA. For example, it may be preferable to isolate only mature, cytoplasmic RNA for some studies on expression profiling. Alternatively it may be desirable to isolate nuclear RNA in order to investigate and study pre-processed (non-spliced) RNA. Furthermore, this kit can be used to isolate RNA for downstream applications where it is necessary to avoid DNA contamination, since the cytoplasmic fraction has been shown to be free of all traces of genomic DNA. 

Details

Supporting Data

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Kit Specifications
Maximum Column Binding Capacity	Up to 50 µg RNA
Maximum Column Loading Volume	650 µL
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Time to Complete 10 Purifications	45 minutes
RNA Yield HeLa (1 x 106) – Cytoplasmic RNA HeLa (1 x 106) – Nuclear RNA	15 µg ≤ 3.5 µg

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.

Component	Cat. 21000 (50 preps)	Cat. 37400 (100 preps)
Lysis Buffer J	20 mL	2 x 20 mL
Buffer SK	40 mL	2 x 40 mL
Wash Solution A	38 mL	2 x 38 mL
Elution Buffer E	6 mL	2 x 6 mL
Mini Spin Columns	50	100
Collection Tubes	50	100
Elution Tubes (1.7 mL)	50	100
Product Insert	1	1

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

Description

Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.

The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit. Details of the target and priming specificity are included in the individual handbooks above.

Packaged, optimised and ready to use. Expect Better Data.

Exceptional value for money
Rapid detection of all clinically relevant subtypes
Positive copy number standard curve for quantification
Highly specific detection profile
High priming efficiency
Broad dynamic detection range (>6 logs)
Sensitive to < 100 copies of target Accurate controls to confirm findings 150 reactions