Overview

• Robust Lysis Solution processes even the most challenging plant species such as pine needle and grape 
• No phenol extractions
• DNA and all sizes of RNA are recovered, including microRNA
• High quality DNA and RNA are purified simultaneously using the same spin column
• No need to split the lysate​
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

This kit provides for rapid spin column isolation and purification of total RNA and genomic DNA simultaneously from a single plant sample without splitting the lysate. Norgen’s plant lysis solution is highly robust and effective over a wide range of plants including challenging samples.  The total RNA and genomic DNA are both column purified in under 30 minutes.   Since RNA and DNA are isolated without splitting the lysate, variability and inconsistent results are reduced.​​ All sizes of RNA including microRNA are recovered without the need for phenol.  Optional on-column DNase and RNase treatments provide flexibility to isolate DNA-free RNA or RNA-free DNA respectively.  Isolated nucleic acids are of a high quality and yield, and are ready for downstream use including PCR, qPCR, RT-PCR, qRT-PCR, sequencing and more. 

Background

It is often necessary to isolate total RNA and genomic DNA from a single plant sample, such as for studies of gene expression, mutant or transgenic plant characterization, and host plant-pathogen characterization. This is of great benefit when isolating RNA and DNA from precious, difficult to obtain or very small samples. Furthermore, gene expression analysis will be more reliable since the RNA and DNA are derived from the same sample, therefore eliminating inconsistent results.

Details

Supporting Data

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Kit Specifications
Column Binding Capacity	50 μg for RNA 15 μg for genomic DNA
Maximum Column Loading Volume	650 μL
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Maximum Amount of Starting Material: Plant Tissues Plant Cells	100 mg 5 x 106
Time to Complete 10 Purifications	30 minutes
Average Yields* Peach Leaves (100 mg)	40 μg RNA, 5 μg gDNA

* Yield will vary depending on the type of sample processed

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. These reagents should remain stable for at least 2 years in their unopened containers.

Component	Cat. 24400 (50 preps)
Lysis Buffer M	40 mL
Binding Buffer I	7 mL
Solution WN	18 mL
Wash Solution A	38 mL
Elution Buffer E	20 mL
Enzyme Incubation Buffer B	6 mL
Filter Columns	50
Spin Columns	50
Collection Tubes	100
Elution Tubes (1.7 mL)	50
Product Insert	1

Introduction

HiPure FFPE Nucleic acid Kit supplies a simple and rapid DNA 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 20 minutes (not including digestion time). DNA can be directly used for downstream applications such as PCR, southern blot and viral DNA detection, etc.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from FFPE tissue samples
Applications	PCR, Southern Blot and viral DNA detection, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples
Sample amount	<20mg
Elution volume	>15µl
Time per run	≤20 minutes
Liquid carrying volume per column	4ml
Binding yield of column	100μg

Principle

Hipure FFPE Nuclear acid kit adopts silica gel column purification. The sample is deparaffinated by xylene and digested by lysate and protease. After decrosslinked at 90 ℃, DNA/RNA is released into the lysate. Adding ethanol to adjust the binding conditions, the sample is transferred to the column where DNA/RNA is adsorbed on the membrane and protein is removed without adsorption. Protein and other impurities are washed by buffer GW1, and the salt is removed by buffer GW2. Finally, the DNA / RNA is eluted by low salt buffer.

Advantages

• Safety – deparaffinating without contacting with xylene or other toxic solution
• Fast – without overnight incubation and digestion, several samples can be extracted within 2hours
• High efficiency – remove the formaldehyde modification of DNA, greatly enhancing the sensitivity of PCR
• High yield – most optimal process to ensure the highest recovery
• High recovery – silica gel column purification method can recover nucleic acid at the level of PG

Kit Contents

Contents	IVD3126
Purification Times	100 Preps
HiPure DNA Mini Columns I	100
2ml Collection Tubes	100
Buffer DPS	70 ml
Buffer ATL	30 ml
Buffer AL	30 ml
Buffer GW1*	44 ml
Buffer GW2*	20 ml
Proteinase K	50 mg
Protease Dissolve Buffer	6 ml
Buffer AE	20 ml

Storage and Stability

Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 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. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

Experiment Data

HiPure FFPE Nucleic acid Kit supplies a simple and rapid DNA 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 20 minutes (not including digestion time). DNA can be directly used for downstream applications such as PCR, southern blot and viral DNA detection, etc.

Bis-propargyl-PEG8 consists of two propargyl groups which can react with azide-bearing compounds or biomolecules in copper catalyzed Click Chemistry to form stable triazole linkage. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Bis-propargyl-PEG8 consists of two propargyl groups which can react with azide-bearing compounds or biomolecules in copper catalyzed Click Chemistry to form stable triazole linkage. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.