4x100ml Low Speed Angle Rotor 

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 1 x 1010 AAV particles as determined by qPCR
AAV Vector Serotype	Any
Average Recovery	> 80%
Input Type	Cells, media, or mixed
Input Volume	8 mL – 45 mL
Minimum Elution Volume	1 mL
Time to Complete 4 Purifications	2 – 2.5 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

Description

The EzRNA™ RNA Capping System is a user-friendly product for post-transcriptional RNA modification. Both Vaccinia Capping Enzyme and 2’-O-Methyltransferase are included in the package, which are able to perform in a single reaction. The Vaccinia Capping Enzyme attach 7-methylguanylate cap (m7Gppp, Cap-0) to the 5′ end of RNA to form m7Gppp5’N-RNA (Cap-0 RNA). The 2′-O-methyltransferase utilizes Cap-0 RNA as a substrate, employing S-adenosine methionine (SAM) as a methyl donor to methylate 2′ -OH of the first nucleotide at the 5′ end of Cap-0 RNA, resulting in the formation of Cap-1 RNA.

Features

• 2’-O-Methyltransferase included for Cap-1 RNA
• High capping efficiency
• High stability
• RNase inhibitor is included to enhance the stability of capping reaction

Application

• Generation of 5’Cap-0 (m7Gppp) and Cap-1 (m7GpppNm-) RNA by enzymatic reaction
• mRNA synthesis for in vitro translation
• Gene expression studies
• mRNA vaccine development and therapeutics

Storage

-20°C for 24 months

The EzRNA™ RNA Capping System is a user-friendly product for post-transcriptional RNA modification. Both Vaccinia Capping Enzyme and 2’-O-Methyltransferase are included in the package, which are able to perform in a single reaction. The Vaccinia Capping Enzyme attach 7-methylguanylate cap (m7Gppp, Cap-0) to the 5′ end of RNA to form m7Gppp5’N-RNA (Cap-0 RNA). The 2′-O-methyltransferase utilizes Cap-0 RNA as a substrate, employing S-adenosine methionine (SAM) as a methyl donor to methylate 2′ -OH of the first nucleotide at the 5′ end of Cap-0 RNA, resulting in the formation of Cap-1 RNA.