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 

ExcelRT™ Reverse Transcription Kit is a complete, efficient and convenient kit to synthesize high quality first strand cDNA. This kit contains ExcelRT™ Reverse Transcriptase, which is able to synthesize the first strand cDNA at 37~50°C. The ExcelRT™ Reverse Transcriptase is a recombinant Moloney Murine Leukemia Virus (M-MLV) reverse transcriptase, which is designed to reduce RNase H activity and create better thermal stability. This kit also contains RNAok™ RNase Inhibitor, which is active against RNase A, RNase B, and RNase C. This product is supplied with oligo (dT)₂₀ and random hexamers, which are used to synthesize cDNA from poly(A) tailed mRNA and total RNA, respectively. 

Features

• Contains all components for reverse transcription 
• High yield 
• Thermostable, up to 50°C, during first strand synthesis 
• High processivity, generating cDNA up to 8 kb 
• Reduced RNase H ribonuclease activity 

Application

• Generation of first strand cDNA from total RNA or mRNA.
• Suitable for generating cDNA from RNA with strong secondary structure which can be reduced at temperature up to 50°C.

Storage

-20°C for 24 months 

High yield

High sensitivity

Thermostable, up to 50°C, during first strand synthesis

 

Contents

ComponentVolume    ExcelRT™ Reverse Transcriptase (200 U/μl) 100 μl      RNase Inhibitor (20 U/μl)100 μl      5X RT Buffer (DTT)500 μl      dNTPs Mix (10 mM each)200 μl      Oligo (dT)20 (50 μM)100 μl      Random Hexamers (100 μM)100 μl      DEPC-Treated H2O  1 ml x 2

Storage Buffer

Reverse Transcriptase: 20 mM Tris-HCl (pH 7.5), 200 mM NaCl, 0.1 mM EDTA, 1 mM DTT, stabilizer and 50% (v/v) glycerol

RNase Inhibitor: 40 mM HEPES-KOH (pH 7.5), 100 mM KCl, 8 mM DTT, 0.1 mM EDTA, stabilizer and 50% (v/v) glycerol

5X RT buffer (DTT)

250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl₂ and 50 mM DTT

Storage

-20°C for 24 months 

ExcelRT™ Reverse Transcription Kit is a complete, efficient and convenient kit to synthesize high quality first strand cDNA. This kit contains ExcelRT™ Reverse Transcriptase, which is able to synthesize the first strand cDNA at 37~50°C. The ExcelRT™ Reverse Transcriptase is a recombinant Moloney Murine Leukemia Virus (M-MLV) reverse transcriptase, which is designed to reduce RNase H activity and create better thermal stability. This kit also contains RNAok™ RNase Inhibitor, which is active against RNase A, RNase B, and RNase C. This product is supplied with oligo (dT)20 and random hexamers, which are used to synthesize cDNA from poly(A) tailed mRNA and total RNA, respectively.

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.