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,

Disulfide Biotin alkyne is an azide-activated cleavable biotin probe that allows for efficient recovery of avidin-bound protein complexes in affinity-based assays. This reagent contains a biotin moiety linked to an alkyne group through a spacer arm containing a cleavable disulfide linker. Under reducing conditions (50 mM dithiothreitol, 10 mM 2-mercaptoethanol or 1% sodium borohydride), the disulfide bonds are cleaved, releasing the biotin tag and any avidin conjugate bound to it.

Disulfide Biotin alkyne is an azide-activated cleavable biotin probe that allows for efficient recovery of avidin-bound protein complexes in affinity-based assays. This reagent contains a biotin moiety linked to an alkyne group through a spacer arm containing a cleavable disulfide linker. Under reducing conditions (50 mM dithiothreitol, 10 mM 2-mercaptoethanol or 1% sodium borohydride), the disulfide bonds are cleaved, releasing the biotin tag and any avidin conjugate bound to it.