Overview

• AAV Purification from any input – cell fraction or media fraction
• High AAV recovery, up to 90%
• No specialized equipment needed
• Purification from a variety of AAV serotypes (including AAV6 and AAV9)
• Yields highly active AAV for in vivo and in vitro experiments
• Purification is based on spin column chromatography that uses Norgen’s resin separation matrix

Recombinant adeno-associated virus (AAV) vectors are highly promising tools for both in vitro and in vivo gene transfer. Norgen’s AAV Purification Kits provide fast and simple procedures for concentrating and purifying AAV vectors from cell lysate and cell culture media. Purification is based on precipitation onto Norgen Biotek’s proprietary resin. Contaminating cellular debris is largely removed from the sample via a centrifugation step, while contaminating DNA and RNA is reduced using enzymatic digestion. AAV vector purified in this manner is highly active for use in in vitro and in vivo transduction experiments.

AAV Purification Kit

Norgen’s AAV Purification Kit contains sufficient materials for 15 preparations (33.5 mL per prep of supernatant (SN) or a total of 500 mL of supernatant input). Approximately 1 mL of cell pellet can be purified per prep, up to a maximum of 15 mL of cell pellet in total for the entire kit. Up to 33X sample concentration.

AAV Purification Mini Kit

Each spin column is able to concentrate and purify AAV from 0.5-8 mL of cell pellet, cell culture media, or cells and culture media mixed together. Up to 50X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (200 µL). Preparation time for 4 samples is 1.5 hours, with 45 minutes of hands-on time.

AAV Purification Midi Kit

Each spin column is able to concentrate and purify AAV from 8 mL up to 45 mL of input consisting of cell pellet, cell culture media, or cells and culture media mixed together. Up to 50X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (1 mL). The kit may be used to purify up to 8 x 25 mL or 4 x 45 mL of samples using the included columns. Preparation time for 4 samples is approximately 2 to 2.5 hours, with 1.5 hours of hands on time.

AAV Purification Maxi Kit (Slurry Format)

Each spin column is able to concentrate and purify AAV from 45 mL to 90 mL of input consisting of cell pellet, cell culture media, or cells and culture media mixed together. Up to 200X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (1-10 mL) using the optional concentration step. The kit may be used to purify up to 1 x 900 mL samples or 10 x 45-90 mL samples using the included columns. Preparation time for 1 x 900 mL sample is approximately 2.5 to 3.5 hours, with an optional concentration step requiring an additional 30 min.

Details

Supporting Data

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Kit Specifications
Column Binding Capacity	At least 5 x 109 AAV particles as determined by qPCR
AAV Vector Serotype	Any
Average Recovery	> 80%
Input Type	Cells, media, or mixed
Input Volume	0.5 mL – 8 mL
Minimum Elution Volume	200 µL
Time to Complete Purifications	1 – 2 hours

 
Storage Conditions and Product Stability
DNAse I and RNAse A should be stored at -20°C upon arrival. Elution Buffer O should be stored tightly capped at 4°C upon arrival. All other solutions should be kept tightly sealed and stored at room temperature. Once opened, the solutions should be stored at 4°C. This kit is stable for 1 year after the date of shipment.

Component	Cat. 66100 (15 preps)	Cat. 63200 (20 preps)	Cat. 63300 (4-8 preps)	Cat. 63250 (1-10 preps)
Lysis Buffer S	5.5 mL	5.5 mL	5.5 mL	20 mL
DNAse I	–	2 x 25 uL	2 x 25 uL	210 μL
RNAse A	–	60 μL	60 μL	240 μL
HL-SAN Nuclease	102 μL	–	–	–
Binding Buffer A	20 mL	4 mL	4 mL	2 x 8 mL
Purification Solution C	60 mL	–	–	–
Purification Solution D	130 mL	–	–	–
Wash Solution C	2 x 130 mL	60 mL	60 mL	3 x 60 mL
Slurry E	12.5 mL	–	–	2 x 14.5 mL
Elution Buffer O	66 mL	8.5 mL	8.5 mL	66 mL
Protein Neutralizer	4 mL	4 mL	4 mL	4 mL
Spin Columns	–	20	–	–
Mini Spin Columns	–	20	–	–
Midi Spin Columns (grey contents) with Collection Tubes	–	–	8	10
Midi Spin Columns (white contents) with Collection Tubes	–	–	8	–
Maxi Spin Columns (grey contents) with Collection Tubes	–	–	–	10
Maxi Spin Columns (white contents) with Collection Tubes	–	–	–	10
Collection Tubes	–	40	–	–
Elution tubes (1.7 mL)	50	20	–	–
Midi Elution tubes (15 mL)	–	–	8	10
Maxi Elution tubes (50 mL)	–	–	–	10
Product Insert	1	1	1	1

PC-Biotin-PEG4-PEG4-alkyne is useful for introducing a biotin moiety to azide-containing biomolecules using Cu(I)-catalyzed Click Chemistry. The extended hydrophilic spacer arm provides better solubility to the labeled molecules in aqueous media. Captured biomolecules can be efficiently photoreleased using near-UV, low intensity lamp (e.g. 365 nm lamp at 1-5 mW/cm2). Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

PC-Biotin-PEG4-PEG4-alkyne is useful for introducing a biotin moiety to azide-containing biomolecules using Cu(I)-catalyzed Click Chemistry. The extended hydrophilic spacer arm provides better solubility to the labeled molecules in aqueous media. Captured biomolecules can be efficiently photoreleased using near-UV, low intensity lamp (e.g. 365 nm lamp at 1-5 mW/cm2). Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Introduction

HiPure DNA Clean Up Kit usesproprietary chemistry and HiPure technology to recover DNA Fragments between20bp-20kbp with yields exceeding 80%. DNA is suitable for ligations, PCR, sequencing,restriction digestion, or various labeling reactions. In addition, this kit canbe also used to recover DNA directly from enzymatic reactions such as PCR andenzyme digestion reactions.

Details

Specifications

Features	Specifications
Main Functions	Recover DNA fragments between 20bp-20kbp from crude DNA and enzymatic reaction solution
Applications	PCR, sequencing, labeling reactions, ligations and restriction digestion, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Crude DNA, enzymatic reaction solution
Sample amount	<150μl
Recovery	80%
Elution volume	≥15μl
Time per run	≤10 minutes
Liquid carrying volume per column	800µl
Binding yield of column	20µg

Principle

The HiPure system uses a simple bind-wash-elute procedure. Gelslices 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 – up to 80% DNA recovery
• Fast – isolation can be completed in 10 minutes by column gel method
• Low elution volume – elution volume can be as low as 10μl
• Efficient – efficient removal of various inhibitory factors

Kit Contents

Contents	D214102	D214103
Purification Times	100 Preps	250 Preps
Buffer GWP	30 ml	60 ml
Buffer DW2	20 ml	50 ml
Elution Buffer	30 ml	60 ml
HiPure DNA Mini Columns I	50	250
2 ml Collection Tubes	50	250

Storage and Stability

HiPure DNA Clean Up Kit components are guaranteed for at least one year when stored at room temperature. If any precipitates form in the buffers, warm at 37℃ to dissolve.

HiPure DNA Clean Up Kit usesproprietary chemistry and HiPure technology to recover DNA Fragments between20bp-20kbp with yields exceeding 80%. DNA is suitable for ligations, PCR, sequencing,restriction digestion, or various labeling reactions. In addition, this kit canbe also used to recover DNA directly from enzymatic reactions such as PCR andenzyme digestion reactions.