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.

Video

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,

6-Formyl-2-pyridine-PEG4-DBCO is a bifunctional PEG linker featuring an aldehyde and a DBCO function. DBCO is a strained cyclooctyne which reacts with azide groups on target molecules while the aldehyde group is a versatile electrophile that is reactive towards amines, hydroxylamines, and hydrazines.

6-Formyl-2-pyridine-PEG4-DBCO is a bifunctional PEG linker featuring an aldehyde and a DBCO function. DBCO is a strained cyclooctyne which reacts with azide groups on target molecules while the aldehyde group is a versatile electrophile that is reactive towards amines, hydroxylamines, and hydrazines.

Product Description

High Sensitivity & Specificity DNA Isothermal Amplification Kit NFO 48 Tests/Kit by Amp-future

Product Detail

Kit Storage and term of Validity

Storage term: stored at ≤-20℃,keep away from light, avoid heavy weight and repeated freezing and thawing.

Term of Validity: 14 months

Isothermal nucleic acid Principle Summary

The kit is based on room and constant temperature nucleic acid rapid amplification technology, its principle is that at room and constant temperature, the recombinase and primer form a protein/single-stranded nucleotide complex Rec/ssDNA, and invade the double-stranded DNA template with the help of auxiliary proteins and single-stranded binding protein SSB; then form a D-loop region at the invasion point and start to scan the DNA duplex, after finding the target region complementary to the primer and disintegration of the complex Rec/ssDNA, the polymerase also binds to the 3′ end of the primer to start the chain extension. The kit relies on the role of NFO enzyme and adds the designed specific molecular probes according to the template, and get the result by colloidal gold technology (sandwich method).

Isothermal nucleic acid Product Features

1/ High sensitivity and specificity, short reaction time.

2/ The reagent form is freeze-dried, stable and easy to operate.

3/ The reaction can be operated by metal bath and water bath pot without purchasing expensive PCR apparatus.

Technical Parameters:

Parameters	Details
Product Name	DNA Isothermal Amplification Kit NFO
Manufacturer	Amp-future
Storage Temperature	-20°C
Kit Components	Enzymes, Buffers ,Reagents
Packaging	48 Tests/box
Detection Limit	500-1000copies/µL
Shipping	ICE
Test Time	5-20mins

Isothermal nucleic acid Applications

Suitable for DNA isothermal rapid amplification kit(NFO type)

Primer: Require pair of nucleotide primers with the length of 25-35 bp.

DNA basic kit reaction temperature is 39 to 42℃ and time is 5-20 minutes.

Notes

1/ Please avoid nucleic acid contamination and set blank control during reaction due to the high sensitivity of the kit.

2/ Please take out the required quantity of MIRA reaction units for the experiment, and put the rest under storage conditions when performing the experiment.