Overview

• Sequentially purify total RNA (and miRNA), DNA and proteins from a single sample
• No sample splitting or need to use phenol or precipitation methods
• Purify RNA/DNA/Protein from cultured animal cells, tissues, blood, bacteria, yeast, fungi or plants
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

RNA/DNA/Protein Purification Plus Kit

This kit provides a rapid method for the high throughput isolation and purification of total RNA, DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, or yeast. The kit employs two columns: 1) for gDNA purification and 2) for RNA purification utilizing Norgen’s resin columns (superior for the binding of all RNA sizes including miRNA). The proteins are also purified on the second column after RNA elution. The proteins are eluted in buffer and are ready for downstream application without any further clean up required. The proteins can be quantified directly, used in western blots, ELISA or mass spectrometry. This kit provides a rapid spin-column method for the isolation and purification of total RNA, genomic DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, yeast, fungi or plants.

RNA/DNA/Protein Purification Plus Micro Kit

This kit provides a rapid spin-column method for the isolation and purification of total RNA, DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, microdissected samples including LCM, stem cells, sorted cells, and CTC. The total RNA, genomic DNA and proteins are all column purified in less than 30 minutes. The RNA and DNA can be eluted in as little as 20 µL while the protein can be eluted in as little as 50 µL. This kit provides the same performance as if the samples were isolated from dedicated kits.

RNA/DNA/Protein Purification 96-Well Plus Kit

The kit employs two plates: 1) for DNA purification and 2) for RNA purification utilizing Norgen’s resin (superior for the binding of all RNA sizes including miRNA). Please see the protocol schematic below.

Details

Supporting Data

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Kit Specifications
Maximum Column Binding Capacity	50 μg for RNA 20 μg for DNA 200 μg for protein
Maximum Column Loading Volume	650 μL
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Size of DNA Purified	≥ 30 kb
Maximum Amount of Starting Material: Animal Cells Animal Tissues Blood Bacteria Yeast Fungi Plant Tissues	5 x 106 cells 25 mg (for selected tissues) 100 μL 1 x 109 cells 1 x 108 cells 50 mg 50 mg
Time to Complete 10 Purifications	30 minutes
Average Yield:HeLa Cells (1 x 106 cells)HeLa Cells (1 x 106 cells)HeLa Cells (1 x 106 cells)	15 μg RNA8 μg DNA150 μg protein

 
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. 47700 (50 preps)	Cat. 51600 (50 preps)	Cat. 51700 (96 preps)
Buffer SKP	40 mL	40 mL	–
Lysis Buffer Q	–	–	40 mL
Wash Solution A	2 x 38 mL 1 x 18 mL	2 x 38 mL 1 x 18 mL	2 x 38 mL 1 x 18 mL
Elution Solution A	6 mL	6 mL	20 mL
Elution Buffer F	15 mL	15 mL	2 x 15 mL
Wash Solution C	30 mL	30 mL	60 mL
Binding Buffer A	8 mL	8 mL	8 mL
Elution Buffer C	8 mL	8 mL	30 mL
Protein Neutralizer	4 mL	4 mL	4 mL
Protein Loading Dye	2 mL	2 mL	3 x 2 mL
gDNA Purification Columns	50	–	–
gDNA Purification Micro Columns	–	50	–
gDNA Purification 96-Well Plate	–	–	1
RNA/Protein Purification Columns	50	–	–
RNA/Protein Purification Micro Columns	–	50	–
RNA/Protein Purification 96-Well Plate	–	–	1
Collection Tubes	150	150	–
Collection Plate	–	–	5
Elution Tubes (1.7 mL)	150	150	–
Elution Plate	–	–	3
Lysis Preparation Plate	–	–	2
Adhesive Tape	–	–	4
Product Insert	1	1	1

Description

Salmonella spp. are members of the family Enterobacteriaceae. They are Gram-negative, facultatively anaerobic, flagellated, rod-shaped organisms. They are approximately 0.7 to 1.5 µm in diameter and 2 to 5 µm in length and responsible for a large number of cases of foodborne illness throughout the world. Salmonella have circular DNA genomes with a mean length of approximately 4530 kb, although this can vary by up 1000 kb. Salmonella classification is extremely complex, however, the genus is divided into two species: S. enterica and S.bongori. S. enterica is then itself divided into 6 biochemically distinct subspecies and the Salmonella genus is further classified into serovars (serotypes) based on the lipopolysaccharide (O), flagella protein (H), and sometimes the capsular (VI) antigens. There are more than 2500 known serovars and within a serovar there may be strains that differ in virulence.

Salmonella are mainly transmitted by the faecal-oral route. They are carried asymptomatically in the intestines or gall bladder of many animals, being continuously or intermittently shed in the faeces. Humans can become infected if they do not wash their hands after contact with infected animals or animal faeces. In such instances the bacteria adhere to and enter the cells of the intestinal epithelium. The toxins produced by the bacteria can damage and kill the cells that line the intestines, which results in intestinal fluid loss. The bacteria can survive for weeks in a dry environment and far longer in water thus they are frequently present in polluted waters. Salmonella can also be carried latently in the mesenteric lymph nodes or tonsils; these bacteria are not shed, but can become reactivated after stress or immunosuppression. In addition, fomites and vectors can spread Salmonella and vertical transmission occurs in birds, with contamination of the vitalize membrane, albumen and possibly the yolk of eggs. Salmonella spp. can also be transmitted in utero in mammals.

There are two different disease conditions that are distinct to salmonellosis; gastroenteritis and enteric typhoid fever. The gastroenteritis is a nonsystemic infection of the intestinal tract and regional lymph nodes that gives rise to headache, muscle aches, diarrhoea, vomiting, abdominal cramping, chills, fever, nausea and dehydration. In contrast, the enteric typhoid fever is a systemic disease in which the microorganism replicates within the cells of the reticuloendothelial system. The symptoms usually appear 6 to 72 hours after ingesting contaminated food although individuals can be infected with the bacteria without having symptoms. Those with and without symptoms shed the bacteria in their stool and it is important that personal hygiene be maintained at all times.

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

Introduction

This kit is suitable for high-throughput extraction of total RNA from paraffin embedded tissue samples, including miRNA. The kit is based on superparamagnetic magnetic particle purification technology. The extraction process does not require the use of toxic phenolic chloroform extraction or time-consuming alcohol precipitation.The whole extraction process only takes 120 minutes. The obtained RNA can be directly used in RT-PCR. The high-throughput extraction can be realized by 96 hole magnetic frame and 96 holeo scillator, various nucleic acid extractors (such as KingFisherFle, MagMix32, MagMix 96) or cooperating with the pipette workstation (Hamilton, Tican) for automatic extraction.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from FFPE tissue
Applications	RT-PCR, cDNA synthesis, second generation sequencing
Products	RNA, miRNA
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Adaptive instrument	Nucleic acid extractor, pipetting workstation
Sample type	FFPE slice, FFPE embedded tissue
Sample amount	≤6 slices

Principle

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Protease. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.

Advantages

• Fast – several samples can be extracted in 120 minutes by column method
• Safe – no phenol chloroform extraction required

Kit Contents

Contents	IVD3022
Purification Times	200 Preps
MagBind Particles	4.5 ml
Proteinase K	100 mg
Protease Dissolve Buffer	6 ml
DNase I	4 x 600 µl
DNase Buffer	30 ml
Buffer DPS	200 ml
Buffer FRL	40 ml
Buffer AL	40 ml
Buffer MW1*	110 ml
Buffer MW2*	2 x 50 ml
Nuclease Free Water	30 ml

Storage and Stability

Proteinase K and MagBind Particles should be stored at 2–8°C upon arrival. DNase I should bestored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K and MagBind 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 theseconditions.

This kit is suitable for high-throughput extraction of total RNA from paraffin embedded tissue samples, including miRNA. The kit is based on superparamagnetic magnetic particle purification technology. The extraction process does not require the use of toxic phenolic chloroform extraction or time-consuming alcohol precipitation.The whole extraction process only takes 120 minutes. The obtained RNA can be directly used in RT-PCR. The high-throughput extraction can be realized by 96 hole magnetic frame and 96 holeo scillator, various nucleic acid extractors (such as KingFisherFle, MagMix32, MagMix 96) or cooperating with the pipette workstation (Hamilton, Tican) for automatic extraction.