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

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

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,

Product Description

Amp-future DNA Isothermal Amplification Kit NFO: Freeze-Dried Reagent

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.

Overview

• Isolate all sizes of circulating and exosomal RNA, including microRNA
• Versatile plasma and serum input volumes
• Concentrate circulating and exosomal RNA into small elution volumes
• Isolate inhibitor-free circulating and exosomal RNA
• Bind and elute all RNA irrespective of size or GC content, without bias
• Available in Spin Column or 96-well plate formats
• Process 96-well plates using vacuum or centrifugation
• Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
• Compatible with Streck Cell-Free RNA BCT® Tubes
• Purification is based on Norgen’s proprietary resin matrix

These kits are able to isolate all sizes of circulating and exosomal RNA, including microRNA, without the use of phenol or chloroform. The slurry format provides an advantage over other available kits in that it does not require extension tubes for the purification of free-circulating and exosomal RNA from large sample volumes. RNA can be isolated from either fresh or frozen samples using this kit. This kit is suitable for the isolation of RNA from serum or plasma prepared from blood collected only on either EDTA or citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications including RT-PCR. The purified plasma/serum free-circulating and exosomal RNA is eluted in an elution solution that is compatible with PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS.

Free-circulating plasma and serum RNA can serve as both tumor- and fetal-specific markers for cancer detection and prenatal diagnosis. As well, free-circulating RNAs have the potential to provide biomarkers for other disease states. Free-circulating RNA in plasma or serum are usually present as short fragments of less than 1000nt, and free-circulating miRNA (21nt) can also be found in plasma and serum.

Plasma/Serum Circulating and Exosomal RNA Purification Kit (Slurry)

Norgen’s Plasma/Serum Circulating and Exosomal RNA Purification Kit (Slurry Format) provides a fast, reliable and simple procedure for isolating circulating RNA and exosomal RNA from plasma and serum samples ranging from 0.25 mL to 5 mL.

Plasma/Serum Circulating and Exosomal RNA Purification Mini Kit (Mini Slurry)

Norgen’s Plasma/Serum Circulating and Exosomal RNA Purification Mini Kit (Slurry Format) provides a fast, reliable and simple procedure for isolating circulating and exosomal RNA from plasma and serum samples ranging from 0.25 mL to 2 mL.

Plasma/Serum Circulating and Exosomal RNA Purification 96-Well Kit (High Throughput Slurry)

Norgen’s Plasma/Serum Circulating and Exosomal RNA Purification 96-Well Kit (Slurry Format) provides a high throughput method for isolating circulating and exosomal RNA from plasma and serum samples using either a vacuum manifold or centrifugation.

Plasma/Serum Circulating and Exosomal RNA Purification Maxi Kit (Maxi Slurry)

Norgen’s Plasma/Serum Circulating and Exosomal RNA Purification Maxi Kit (Slurry Format) provides a fast, reliable and simple procedure for isolating circulating RNA and exosomal RNA from various amounts of plasma/serum ranging from 2 mL to 5 mL.

Details

Supporting Data

Figure 1 / 5

Previous

Next

Click for expanded view

Kit Specifications – 96-well
Minimum Plasma/Serum Input	0.25 mL
Maximum Plasma/Serum Input	2 mL
Size of RNA Purified	All sizes, including microRNA
Time to Complete Purification	< 1 hour

Cat.29500 Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.

Component	Cat. 51000 (50 preps)	Cat. 29500 (96 preps)	Cat. 42800 (50 preps)	Cat. 50900 (25 preps)
Slurry C1	–	–	–	6 mL
Slurry C2	12 mL	–	12 mL	–
Slurry C3	–	20 mL	–	–
Lysis Buffer A	2 x 130 mL	4 x 100 mL	2 x 130 mL	2 x 130 mL
Wash Solution A	38 mL	2 x 38 mL	38 mL	18 mL
Elution Solution A	6 mL	20 mL	6 mL	6 mL
Mini Filter Spin Columns	50	–	50	25
96-Well Filter Plate	–	1	–	–
Adhesive Tape	–	1	–	–
Collection Tubes	50	–	50	25
96-Well Collection Plate	–	1	–	–
Elution Tubes (1.7 mL)	50	–	50	25
96-Well Elution Plate	–	1	–	–
Product Insert	1	1	1	1