GeneAb™ FSH

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

For scaling up and fully automating end-to-end NGS library prep

With the Flex NGS Workstation, you can automate your NGS workflow at the scale you need to increase efficiency, reduce errors, and save hands-on time. The Flex enables you to automate your library prep and pre-sequencing workflows using any leading reagent system on the market, including fragmentation and tagmentation protocols.

Optional add-ons can be purchased at a 10% discount when ordered with the Flex NGS Workstation*

For scaling up and fully automating end-to-end NGS library prep

With the Flex NGS Workstation, you can automate your NGS workflow at the scale you need to increase efficiency, reduce errors, and save hands-on time. The Flex enables you to automate your library prep and pre-sequencing workflows using any leading reagent system on the market, including fragmentation and tagmentation protocols.

Optional add-ons can be purchased at a 10% discount when ordered with the Flex NGS Workstation*

The dsDNA Quantification Broad Range Kit and dsDNA Quantification High Sensitivity Kit are developed for double stranded DNA quantification using microplate readers. The DNA Quantification BR kit includes BR Dye, BR Dilution Buffer, and BR DNA Standards. The DNA Quantification HS kit includes HS Dye, HS Dilution Buffer, and HS DNA Standards. Simply dilute the Dye with the Dilution Buffer, add DNA sample, then read the concentration using microplate reader. The BR assay is accurate in the linear range from 4 to 1000 ng and the HS assay is accurate in the linear range from 0.2 to 100 ng. Both kits are highly selective for double-stranded DNA over RNA.

The Quantification Kits have several advantages over traditional DNA quantitation such as sensitivity, stability, and tolerance of contaminants.

Both kits reduce the cost of DNA quantification by 60% as compared to other brands.

Selectivity and sensitivity of the kits

Save 60% of the costs.

The performance of the kit is nearly identical to that of Thermo Fisher’s kit (figure below).

Comparison of BioDynami dsDNA Quantification Broad Range Kit with Thermo Fisher kit.

Comparison of BioDynami dsDNA Quantification High Sensitivity Kit with Thermo Fisher kit.

Common contaminants such as salts, free nucleotides, solvents, detergents, RNA, single stranded DNA, or protein are well tolerated in the assay (Table 1).