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
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
IVD3182
Purification Times
50
Buffer ACL
250 ml
Buffer ACB*
300 ml
Buffer DCW1*
22 ml
Buffer DCW2*
10 ml
Proteinase K
540 mg
Protease Dissolve Buffer
30 ml
Carrier RNA
110 μg
Nuclease Free Water
20 ml
HiPure CFDNA Mini Columns
50
2 ml Collection Tubes
100
Extender Tube
50
Vac-Connector
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
Document
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,
HiPure Gel Pure DNA Kit uses proprietary chemistry and HiPure technology to recover DNA fragments between 60bp-10kbp with yields exceeding 80%. DNA is suitable for ligations, PCR, sequencing, restriction digestion, or various labeling reactions. In addition, this kit can be also used to recover DNA directly from enzymatic reactions such as PCR and enzyme digestion reactions.
Details
Specifications
Features
Specifications
Main Functions
Recover DNA fragments >100bp from agarose gel(<0.5g), purification of DNA from PCR, enzymatic reaction solution or crude gDNA
Applications
PCR, NGS, labeling, ligation and enzyme digestion, etc.
Purification method
Mini spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Agarose gel, PCR products, enzyme products
Sample amount
Agarose gel: ≤500mg
Recovery
≥80%
Elution volume
≥15μl
Time per run
≤20 minutes（1-24 samples）
Liquid carrying volume per column
800µl
Binding yield of column
35µg
Principle
The HiPure system uses a simple bind-wash-elute procedure. Gel slices are dissolved in a buffer containing a pH indicator, allowing easy determination of the optimal pH for DNA binding, and the mixture is applied to the column. Nucleic acids adsorb to the silica-gel membrane in the high-salt conditions provided by the buffer. Impurities are washed away and pure DNA is eluted with a small volume of low-salt buffer provided or water, ready to use in subsequent applications.
Advantages
High recovery efficiency – ≥80% DNA recovery
General – recover DNA from gel or enzyme-driven reaction solutions such as PCR
Fast – isolation can be completed in 10-15 minutes by column gel method
Great cost-effectiveness performance
Kit Contents
Contents
D211102
D211103
Purification Times
100 Preps
250 Preps
Buffer GDP
120 ml
250 ml
Buffer DW2
50 ml
2 x 50 ml
Elution Buffer
20 ml
30 ml
HiPure DNA Mini Columns II
100
250
2 ml Collection Tubes
100
250
Storage and Stability
The Kit should be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve.
Experiment Data
Document
HiPure Gel Pure DNA Kit uses proprietary chemistry and HiPure technology to recover DNA fragments between 60bp-10kbp with yields exceeding 80%. DNA is suitable for ligations, PCR, sequencing, restriction digestion, or various labeling reactions. In addition, this kit can be also used to recover DNA directly from enzymatic reactions such as PCR and enzyme digestion reactions.
