Introduction

HiPure DNA Clean Up 96 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 between 60bp-10kbp from PCR, Reactions, crude DNA using 96 well Bind Plate
Applications	PCR, sequencing, labeling reactions, ligations and restriction digestion, etc.
Purification method	96 well Bind Plate
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	PCR product, enzymatic reaction
Sample amount	Appropriate
Recovery	≥80%
Elution volume	≥75μl
Time per run	≤60 minutes
Liquid carrying volume per column	800µl
Binding yield of column	20µg

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
• Fast – isolation can be completed in 60 minutes

Kit Contents

Contents	D212201	D212202	D212203
Purification Times	1 x 96 Preps	4 x 96 Preps	20 x 96 Preps
Buffer GDP	120 ml	400 ml	4 x 400 ml
Buffer DW2	20 ml	100 ml	3 x 100 ml
Elution Buffer	20 ml	20 ml	30 ml
HiPure DNA Plate	1	4	20
2.2ml Collection Plate	1	4	20
1.6 ml Collection Plate	1	4	20
0.8 ml Collection Plate	1	4	20

Storage and Stability

The Kit can 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.

HiPure DNA Clean Up 96 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.

Propargyl-PEG5-alcohol enables Click Chemistry reactions with azide compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Propargyl-PEG5-alcohol enables Click Chemistry reactions with azide compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

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
Resin Binding Capacity (total per kit)	At least 5 x 1010 AAV particles as determined by qPCR
AAV Vector Serotype	AAV6, AAV9 and others
Input Type	Cells, media
Input Volume (AAV supernatant)	1 – 33.5 mL SN per prep (500 mL SN in total)
Input Volume (AAV cell pellet)	1 mL cell pellet per prep (15 mL in total)
Time to Complete Purifications	2.5 to 4.5 hours with 1 hour hands on time
In vivo transduction	Yes

Storage Conditions and Product Stability

HL-SAN Nuclease 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 2 years 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