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
Applications	RT-PCR, cDNA synthesis, second generation sequencing
Products	RNA, miRNA
Purification method	Mini spin column
Purification technology	Silica technology, DNase
Process method	Manual (centrifugation or vacuum)
Sample type	FFPE slice, FFPE embedded tissue
Sample amount	Tissue: <6 mgFFPE: <6 slices
Yield	1-20μg
Elution volume	≥15μl
Time per run	≤30 minutes (after digestion)
Liquid carrying volume per column	800μl

Principle

This product is based on silica column purification. Remove paraffin by Buffer DPS. Sample lysis withproteinase 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 wasfinally eluted with low-salt buffer.

Advantages

• Efficient removal of DNA – unique genomic DNA removal column without DNase treatment
• 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

Kit Contents

Contents	IVD4144
Purification Times	50 Preps
HiPure RNA Micro Columns	50
2ml Collection Tubes	50
Proteinase K	24 mg
Protease Dissolve Buffer	1.8 ml
DNase I	600 μl
DNase Booster Buffer	1.5 ml
Buffer DPS	60 ml
Buffer FRL	15 ml
Buffer RLC	15 ml
Buffer RWC*	10 ml
Buffer RW2*	20 ml
RNase Free Water	10 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 roomtemperature (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.

Description 

The ExcelTaq™ 5X/2X PCR Master Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as a master mix for virtually all PCR applications. The mixture contains all components for PCR with the exception of templates and primers. This not only saves valuable time in the laboratory, but also reduces pipetting and reagent handling errors. The PCR Master Mix is supplied as a 5X/2X concentrated ready-to-use mixture of recombinant Taq DNA Polymerase, reaction buffer, MgCl₂ (TP1120 contains MgSO4), dNTP, and enzyme stabilizer enabling efficient amplification of template in PCR and allows the user to prepare a PCR reagent conveniently. This product is supplied with 6X DNA Loading Dye (Blue) containing two tracking dyes (Xylene cyanol FF and Bromophenol blue) for post PCR analysis through the use of agarose gel electrophoresis.

Features

• 5’→3’ DNA polymerase activity
• No detectable 3’→5′ exonuclease (proofreading) activity
• Generates PCR products with 3′-dA overhangs
• High yield PCR 
• High reproducibility
• Reduced pipetting errors

Applications 

• Routine PCR
• Colony PCR
• High throughput PCR
• Amplification of DNA fragments up to 8 kb
• Generation of PCR products for TA cloning
• DNA labeling

Storage

4°C for 6 months
-20°C for 24 months

The ExcelTaq™ 5X/2X PCR Master Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as a master mix for virtually all PCR applications. The mixture contains all components for PCR with the exception of templates and primers. This not only saves valuable time in the laboratory, but also reduces pipetting and reagent handling errors. The PCR Master Mix is supplied as a 5X/2X concentrated ready-to-use mixture of recombinant Taq DNA Polymerase, reaction buffer, MgCl2 (TP1120 contains MgSO4), dNTP, and enzyme stabilizer enabling efficient amplification of template in PCR and allows the user to prepare a PCR reagent conveniently. This product is supplied with 6X DNA Loading Dye (Blue) containing two tracking dyes (Xylene cyanol FF and Bromophenol blue) for post PCR analysis through the use of agarose gel electrophoresis.