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.

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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,

Overview

• Isolate high quality, high yield plasmid DNA
• Plasmid DNA is ready for various downstream applications including restriction digestion, bacterial transformation, sequencing and more
• Available in 4 formats: MiniPrep, MiniPrep (Magnetic Bead System), 96-Well MiniPrep (Magnetic Bead System), and MaxiPrep

These kits are designed for the rapid preparation of plasmid DNA from Escherichia coli.

Plasmid MiniPrep Kit

This kit is designed for the rapid preparation of plasmid DNA from small cultures of Escherichia coli using convenient spin columns. The plasmid DNA is preferentially purified from other cellular components such as genomic DNA and RNA. This kit is able to purify plasmids up to 13,000 bp in size, and the typical purification yield is up to 20 μg from 1.5 mL of bacterial culture. Purified DNA is of excellent quality for transformation, restriction enzyme digestion, sequencing and more. Also available in a 96-well format.

Plasmid MiniPrep Kit (Magnetic Bead System and High Throughput Magnetic Bead System)

Norgen’s Plasmid MiniPrep Kit (Magnetic Bead System) is designed for the rapid preparation of plasmid DNA from small batch cultures of Escherichia coli. Norgen’s magnetic beads bind DNA under optimized salt concentrations and release the bound DNA under low salt and slightly alkali conditions. The plasmid DNA is preferentially purified from other cellular components such as genomic DNA and RNA. The purified plasmids are fully digestible with all restriction enzymes tested, and are completely compatible with real-time PCR and NGS.

Norgen’s Plasmid MiniPrep Kit (Magnetic Bead System) is also available in a 96-well (HT) format for high throughput applications. Purification with the 96-well plates can be integrated with a robotic automation system.

Plasmid MaxiPrep Kit

This kit is designed for the rapid spin column preparation of plasmid DNA from up to 100 mL of Escherichia coli cultures. The kit allows for the isolation of plasmid DNA with final endotoxin levels of 0.1 EU/µg of DNA or less. The kit is able to purify plasmids up to 13,000 bp in size, and typical yields from a 100 mL culture for a high copy number plasmid are between 0.4 and 1.0 mg. The purified DNA is fully digestible with all restriction enzymes tested, and is completely compatible with manual or automated sequencing to achieve 95-100% accuracy.

Details

Supporting Data

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Kit Specifications
Column Binding Capacity	25 µg
Size of Plasmids Purified	Up to 13,000 bp
Average Yield from 1.5 mL of Culture	Up to 20 µg
Time to Complete 10 Purifications	30 minutes

Storage Conditions and Product Stability

All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the of shipment. The RNase should be stored at -20°C upon arrival. The Resuspension Solution AZ should be stored at 4oC upon addition of RNase enzyme.

Component	Cat. 13300 (50 preps)	Cat. 46400 (250 preps)	Cat. 46500 (4 preps)	Cat. 46600 (20 preps)	Cat. 60300 (50 preps)	Cat. 63000 (192 preps)
Resuspension Solution AZ	12 mL	60 mL	20 mL	100 mL	12 mL	2 x 20 mL
Lysis Buffer N	40 mL	80 mL	40 mL	2 x 80 mL	40 mL	2 x 40 mL
Buffer TN	20 mL	130 mL	55 mL	2 x 130 mL	20 mL	1 x 55 mL 1 x 20 mL
Wash Solution E	12 mL	2 x 18 mL	–	–	–	–
Elution Buffer K	8 mL	30 mL	–	–	8 mL	2 x 8 mL
Wash Solution J	–	–	25 mL	3 x 25 mL	–	–
Elution Buffer J	–	–	24 mL	120 mL	–	–
RNase A	1 vial	1 vial	1 vial	1 vial	1 vial	1 vial
Magnetic Bead Suspension	–	–	–	–	1 x 1.1 mL	4 x 1.1 mL
Spin Columns	50	250	–	–	–	–
Collection Tubes	50	250	–	–	–	–
DNA Maxi Spin Columns with Collection Tubes (Clear ring in column)	–	–	4	20	–	–
Maxi Spin Filter Columns with Collection Tubes (Grey ring in column)	–	–	4	20	–	–
96-Well Plate	–	–	–	–	–	2
Elution Tubes (1.7 mL)	50	250	–	–	50	–
Elution Tubes (50 mL)	–	–	4	20
96-Well Elution Plate	–	–	–	–	–	2
Adhesive Tape	–	–	–	–	–	2
Product Insert	1	1	1	1	1	1

Permagen’s most universal ring magnet plate. From Low elution PCR applications up to 2 mL deep well, the X96 has you covered. No need to purchase two separate plates anymore. Smaller inner ring magnet allows for volumes as low as 5 µL (from PCR plates), larger outer magnet handles up to 2 mL deep well assays

ANSI/ SBS Footprint (127.75mm x 85.50mm) to fit into any automated liquid handling robot on bottom, SBS/ SLAS fit on top to accept any microplate

Integrated Cushion base for maximum recovery. Helps aid in set-up, robot positioning inconsistencies, and labware consumable differences 

Features include solid aluminum alloy construction and hard coat anodized finish for years of trouble-free use, and compatible with any magnetic beads

SKU #

A011623

Features

A011623

Compatibility

• Round bottom plates 
• Pyramid bottom plates 
• Flat Bottom plates
• Deep well plates
• Full-, Semi-, and un-Skirted PCR Plates
• Any magnetic beads
• Any common liquid handlers i.e. Tecan®, Opentrons®, Hamilton®, Agilent®, Beckman®, etc.

Volumes

Maximum – 2 mL (deep well plate

Minimum – 5 µL (pcr plate, or tubes with kit)

Permagen’s most universal ring magnet plate. From Low elution PCR applications up to 2 mL deep well, the X96 has you covered. No need to purchase two separate plates anymore. Smaller inner ring magnet allows for volumes as low as 5 µL (from PCR plates), larger outer magnet handles up to 2 mL deep well assays