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.
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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 |
HiPure Insect DNA Kits provides a simple and rapid solution for total DNA extraction of insect tissue samples. This kit is based on silica gel column purification technology without toxic phenol chloroform extraction and time-consuming alcohol precipitation. The whole extraction process only takes 30 minutes. HiPure Insect DNA Kit can process tissue samples less than 10mg at a time. Hipure Insect DNA 96 kit can process 96 insect tissue samples at a high throughput. The obtained DNA can be directly used in PCR, Southern blot, viral DNA detection and other experiments.
Specifications
| Features | Specifications |
| Main Functions | Isolation total DNA from <10 mg insect tissue |
| Applications | PCR, southern bolt and virus detection, etc |
| Purification method | Midi spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Insect tissue samples |
| Sample amount | <10 mg |
| Elution volume | ≥15μl |
| Time per run | ≤30 minutes |
| Liquid carrying volume per column | 800μl |
| Binding yield of column | 100μg |
Principles
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 (10mm Tris, pH9.0, 0.5mm EDTA).
| Contents | D312902 | D312903 |
| Purification Times | 50 | 250 |
| HiPure DNA Mini Columns I | 50 | 250 |
| 2ml Collection Tubes | 50 | 250 |
| Buffer ITL | 30 ml | 120 ml |
| Buffer IL* | 30 ml | 120 ml |
| Buffer GW1* | 22 ml | 110 ml |
| Buffer GW2* | 20 ml | 2 x 50 ml |
| Proteinase K | 24 mg | 120 mg |
| Protease Dissolve Buffer | 1.8 ml | 15 ml |
| Buffer AE | 15 ml | 60 ml |
Proteinase K 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 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 thiscase buffers should be redissolved before use. Make sure that all buffers areat room temperature when used.
HiPure Insect DNA Kits provides a simple and rapid solution for total DNA extraction of insect tissue samples. This kit is based on silica gel column purification technology without toxic phenol chloroform extraction and time-consuming alcohol precipitation. The whole extraction process only takes 30 minutes. HiPure Insect DNA Kit can process tissue samples less than 10mg at a time. Hipure Insect DNA 96 kit can process 96 insect tissue samples at a high throughput. The obtained DNA can be directly used in PCR, Southern blot, viral DNA detection and other experiments.
