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 Midi Kit enables efficient purification of these circulating nucleic acids from human plasma, serum, or urine.

This product is for research use only.

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Details

Specifications

Features	Specifications
Main Functions	Isolation circulating DNA from 1-5ml plasma, serum, body fluids using vacuum protocol
Applications	qPCR, liquid or solid chip analysis, hybridization and SNP detection, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (vacuum)
Sample type	Serum, plasma and other cell-free fluid samples
Sample amount	1-5ml
Elution volume	≥50μl
Time per run	≤60 minutes
Liquid carrying volume per column	4ml
Binding yield of column	1mg

Principle

This product is based on silica Column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption 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.

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 thelevel of PG by silica gel column purification

Kit Contents

Contents	D318201C	D318202C
Purification Times	10	50
Buffer ACL	50 ml	250 ml
Buffer ACB*	60 ml	300 ml
Buffer DCW1*	4.4 ml	22 ml
Buffer DCW2*	5 ml	10 ml
Proteinase K	110 ml	540 mg
Protease Dissolve Buffer	10 ml	30 ml
Carrier RNA	110 μg	110 μg
Nuclease Free Water	10 ml	20 ml
HiPure CFDNA Mini Columns	10	50
2 ml Collection Tubes	20	100
Extender Tube	10	50
Vac-Connector	10	50

Storage and Stability

Proteinase K, 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 remaining kit 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 should be redissolved before use. Make sure that all buffers are at room temperature when used.

Experiment Data

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

This product provides fast and easy methods for purification of total DNA for reliable PCR and Southern blotting. Total DNA(e.g., genomic, viral, mitochondrial) can be purified from small volume of blood, tissue and dry blood spots.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from 1-10μl blood, <10mg tissue, urine, blood stain, seminal stain
Applications	PCR, southern bolt and virus detection, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Animal tissues, blood stain, urine, seminal stain and various forensic samples
Sample amount	Blood:1-100μl, Tissue:<10mg
Elution volume
Time per run
Liquid carrying volume per column
Binding yield of column

Principle

This product is based on silica Column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption 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 (10mmTris, pH9.0, 0.5mm EDTA).

Advantages

• Fast – several samples can be extracted in 20 minutes (after digestion)
• High purity – purified DNA can be directly used in various downstream applications
• High recovery – DNA can be recovered at the level of PG
• Good repeatability – silica technology can obtain ideal results every time

Kit Contents

Contents	D312502	D312503
Purification Times	50 Preps	250 Preps
Buffer ATL	15 ml	60 ml
Buffer AL	15 ml	60 ml
Buffer GW1*	22 ml	66 ml
Buffer GW2*	20 ml	2 x 50 ml
Carrier RNA	310 μg	2 x 310 µg
Proteinase K	24 mg	120 mg
Protease Dissolve Buffer	1.8 ml	10 ml
Buffer AE	15 ml	60 ml
HiPure DNA Mini Columns I	50	2 x 125
2 ml Collection Tubes	100	5 x 100

Storage and Stability

Carrier RNA and 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 its performance. The remaining kit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under theseconditions.

Experiment Data

This product provides fast and easy methods for purification of total DNA for reliable PCR and Southern blotting. Total DNA(e.g., genomic, viral, mitochondrial) can be purified from small volume of blood, tissue and dry blood spots.

Description

The oasig freeze drying process stabilises all of the active components allowing these reagents to be shipped and stored at room temperature. This hugely simplifies the logistics of purchasing, shipping and using the technology. Whether you are in a sophisticated laboratory in Texas or a mobile field hospital in Timbuktu we can supply complete qPCR kit and reagent packages to your door quickly and cheaply via standard shipping methods without the need for dry ice or a cold chain of any sort.

The performance of the reagents is second to none. We are confident that you will find excellent data quality and even see an improvement in data quality versus many traditional frozen master mixes.

No cold shipping required.  Our unique, oasig Lyophilised qPCR Master Mix in a 150 reaction pack. Perfect to accompany any genesig kit for a DNA target.

Primerdesign real-time PCR reagents are manufactured to the highest standards within our ISO9001:2008 and ISO13485:2012 certified quality management laboratory environment.