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. The extracted products can be used for clinical in vitro detection.

Details

Specifications

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

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,

N-(Propargyl-PEG2)-PEG3-N-Boc is a branched PEG linker with an alkyne group and a t-butyl ester. The alkyne group reacts with azide-bearing compound in coppper catalyzed azide-alkyne Click Chemistry reaction. The Boc group can be deprotected under acidic conditions. The amino group is reactive with carboxylic acids, activated NHS esters, and carbonyls. Reagent grade, for research purpose.

N-(Propargyl-PEG2)-PEG3-N-Boc is a branched PEG linker with an alkyne group and a t-butyl ester. The alkyne group reacts with azide-bearing compound in coppper catalyzed azide-alkyne Click Chemistry reaction. The Boc group can be deprotected under acidic conditions. The amino group is reactive with carboxylic acids, activated NHS esters, and carbonyls. Reagent grade, for research purpose.

Introduction

This product is designed for purification of high-molecular-weight genomic or mitochondrial DNA from a variety of tissue and culture cells. The convenient, scalable purification procedure removes contaminants and enzyme inhibitors such as proteins and divalent cation, and purified DNA is ready for immediate use in sensitive downstream applications or for archiving.

Details

Specifications

Principles

Cells are lysed with ananionic detergent in the presence of a DNA stabilizer. The DNA stabilizer limits the activity of intracellular DNases and also DNases found elsewhere in the environment. RNA is then removed by treatment with an RNA digesting enzyme. Other contaminants, such as proteins, are removed by salt precipitation. Finally, the genomic DNA is recovered by precipitation with alcohol and dissolved in Buffer TE. Purified DNA typically has an A260/A280 ratio between 1.7 and 1.9, and is up to 200 kb in size. The DNA can be safely stored at 2-8°C, -20°C, or -80°C.

Advantages

• Safe – without phenol chloroform extraction
• Environment friendly – the reagents used are safe, non-toxic and without pollution
• High molecular weight – the molecular weight of genomic DNA is about 50-150kb
• High purity – the purified DNA has A260/280=1.8-1.9, A260/230=2.0-2.5
• Unlimited sample size – solution type operation, sample volume can be adjusted at will
• Cost performance – the most economical nucleic acid extraction technology

Kit Contents

Storage and Stability

RNase Solution should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 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.

This product is designed for purification of high-molecular-weight genomic or mitochondrial DNA from a variety of tissue and culture cells. The convenient, scalable purification procedure removes contaminants and enzyme inhibitors such as proteins and divalent cation, and purified DNA is ready for immediate use in sensitive downstream applications or for archiving.