Introduction

Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments, <1000bp (DNA). HiPure Circulating DNA Mini Kit enables efficient purification of these circulating nucleic acids from human plasma, serum, or urine. Samples can be fresh or frozen (provided that they have not been frozen and thawed more than once).

Details

Specifications

Features	Specifications
Main Functions	Isolation circulating DNA from 1ml plasma, serum, body fluids
Applications	qPCR, liquid or solid chip analysis, hybridization and SNP detection, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Serum, plasma and other cell-free fluid samples
Sample amount	1ml
Elution volume	≥30μl
Time per run	≤50 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

This product is based on silica gel purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption plate and filter column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10 Mm Tris,pH 8.0).

Advantages

• High yield – most optimal process, free DNA (>50bp) can be obtained to the maximum extent
• High concentration – low elution volume, ensuring high nucleic acid concentration
• High purity – low alcohol binding method, completely removing inhibitor and protein pollution
• High recovery – DNA can be recovered at the level of PG by silica gel column purification

Kit Contents

Contents	D318102	D318103
Purification Times	50 Preps	250 Preps
Buffer ACL	50 ml	250 ml
Buffer ACB*	60 ml	300 ml
Buffer DCW1*	22 ml	88 ml
Buffer DCW2*	10 ml	50 ml
Proteinase K	120 mg	540 mg
Protease Dissolve Buffer	10 ml	30 ml
Carrier RNA	110 μg	310 μg
Nuclease Free Water	10 ml	30 ml
HiPure CFDNA Mini Columns	50	250
2 ml Collection Tubes	100	500

Storage and Stability

Proteinase K and carrier RNA 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 remainingkit components can be stored dry 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 shouldbe redissolved before use. Make sure that all buffers are at room temperature when used.

Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments,

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.