Introduction

HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from FFPE tissue and section samples (with DNase)
Applications	RT-PCR, quantitative RT-PCR, Northern hybridization, Poly A purification, nucleic acid protection and in vitro translation
Purification method	Mini spin column
Purification technology	Silica technology, DNase
Process method	Manual (centrifugation or vacuum)
Sample type	FFPE tissue sample
Sample amount	6mg
Yield	20μg
Elution volume	≥10μl
Time per run	≤60 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

This product is based on silica column purification. Remove paraffin by Buffer DPS. Sample lysis with proteinase K digestion requires only 15 minutes. After lysis, samples are incubated at 80ºC for 15 minutes. Transfer to an adsorption column and RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, RNA was finally eluted with low-salt buffer.

Advantages

• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Fast – several samples can be extracted in 60 minutes by column method
• Safe – no phenol chloroform extraction required
• Sensitive – RNA can be recovered at the level of PG
• Efficient DNA removal – unique method to effectively remove genomic DNA

Kit Contents

Contents	R414402	D414403
Purification Times	50 Preps	250 Preps
HiPure RNA Micro Columns	50	250
2ml Collection Tubes	50	250
Buffer DPS	60 ml	250 ml
Buffer FRL	15 ml	60 ml
Buffer RLC	15 ml	60 ml
Buffer RWC*	10 ml	50 ml
Buffer RW2*	20 ml	2 x 50 ml
DNase I	600 µl	5 x 600 µl
DNase Booster Buffer	1.5 ml	6 ml
Protease Dissolve Buffer	1.8 ml	10 ml
Proteinase K	24 mg	120 mg
RNase Free Water	10 ml	20 ml

Storage and Stability

Proteinase K 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, Proteinase K 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.

Experiment Data

HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.

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	35 μg for RNA 10 μg for DNA 100 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
Time to Complete 10 Purifications	30 minutes
Maximum Amount of Starting Material:Animal CellsAnimal TissuesBloodPlant / Fungi Tissues	5 x 105 cells5 mg (for selected tissues)50 µL25 mg
Average Yield* HeLa Cells (5 x 105 cells) HeLa Cells (5 x 105 cells) HeLa Cells (5 x 105 cells)	RNA: 7.5 μg DNA: 1-2 μg Protein: 70 μg

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

DBCO-PEG12-acid is an analog of DBCO-Acid with hydrophilic PEG linker and a DBCO group. The DBCO groups is commonly used for Click Chemistry reactions. The hydrophilic PEG chain allows for increased water solubility of compounds in aqueous media. The terminal carboxylic acid can react with primary amine groups in the presence of activators (e.g. EDC, or HATU) to form a stable amide bond. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

DBCO-PEG12-acid is an analog of DBCO-Acid with hydrophilic PEG linker and a DBCO group. The DBCO groups is commonly used for Click Chemistry reactions. The hydrophilic PEG chain allows for increased water solubility of compounds in aqueous media. The terminal carboxylic acid can react with primary amine groups in the presence of activators (e.g. EDC, or HATU) to form a stable amide bond. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.