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 de crosslinked 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

Kit Contents

Contents	D312602	D312603
Purification Times	50 Preps	250 Preps
HiPure DNA Mini Columns I	50	250
2ml Collection Tubes	50	250
Buffer DPS	30 ml	150 ml
Buffer ATL	15 ml	60 ml
Buffer AL	15 ml	60 ml
Buffer GW1*	22 ml	88 ml
Buffer GW2*	12 ml	50 ml
Proteinase K	24 mg	120 mg
Protease Dissolve Buffer	1.8 ml	10 ml
Buffer AE	10 ml	30 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.

Description

Specifications

Clone	IHC019
Source	Mouse Monoclonal
Positive Control	Bladder, Colon Carcinoma, Colon, Thyroid Carcinoma
Dilution Range	1:200

Cytokeratin 19 (CK19) forms intermediate filaments found in the intracytoplasmic cytoskeleton of epithelial tissue and provides mechanical support. Anti-Cytokeratin 19 stains epithelia and epithelial malignancies such as carcinomas of the colon, stomach, pancreas, biliary tract, liver, and breast. Cytokeratin 19 is a useful marker for distinguishing hepatocellular carcinoma from intrahepatic cholangiocarcinoma. This differentiation is improved when stained in combination with Cytokeratin 7, CAM5.2l, Ber-EP4/MOC31, HepPar1 and TTF1. Cytokeratin 19 staining can also be used to recognize thyroid papillary carcinomas.

Overview

• Isolate high quality DNA from a broad variety of phage strains
• High yields of total DNA
• Fast and easy processing using a rapid spin-column format
• No phenol or chloroform extractions or cesium chloride banding required
• High yields of DNA recovered 3-15 µg DNA from 10⁶-10¹⁰ pfu/ mL of enriched phages

This kit provides a rapid spin column method for the purification of total DNA from a broad spectrum of bacteriophages propagated in bacteria grown in liquid cultures. The DNA is isolated without the use of phenol, chloroform or cesium chloride banding procedures. The spin-column based procedure is rapid and can be completed in less than 45 minutes. The kit is highly efficient for processing small volumes of phage supernatant (500 µL – 1 mL) and with the optional DNase and Proteinase K treatments phage DNA yields are maximized while host DNA contamination is minimized. Purified total phage DNA is of the highest integrity, and can be used in a number of downstream applications including PCR, qPCR, Restriction Fragment Length Polymorphism (RFLP), sequencing, cloning, Southern Blot and more.

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Details

Supporting Data

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Kit Specifications
Column Binding Capacity	50 µg
Maximum Column Loading Volume	650 µL
Size of DNA Purified	All sizes
Maximum Amount of Starting Material	1 x 1010 pfu/mL enriched phages
Average Yield*	3-15 µg DNA from 106-1010 pfu/mL of enriched phages
Time to Complete 10 Purifications	45 minutes

* Average yields will vary depending upon a number conditions used and developmental stage.

Storage Conditions and Product Stability
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. 46800 (50 preps)	Cat. 46850 (100 preps)
Lysis Buffer B	40 mL	2 x 40 mL
Wash Solution A	38 mL	2 x 38 mL
Elution Buffer B	8 mL	2 x 8 mL
Spin Columns	50	100
Collection Tubes	50	100
Elution Tubes (1.7 mL)	50	100
Product Insert	1	1