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 | |
| 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 |
TG16E Technical Parameter：
| Max. Speed | 16000rpm |
| Max. RCF | 19040×g |
| Max. Capacity | 6×10ml |
| Time Range | 0~99min |
| RPM/RCF Convert | Yes |
| Noise (dB) | ≤ 65 |
| Acc/Dec | 10 Kinds |
| Speed Accuracy | ±20r/min |
| Voltage(V/Hz) | AC 220V/110V 50HZ/60HZ |
| Size (W x D x Hmm) | 370×290×215mm |
| Net Weight(Kg) | 16KG |
| Certificates | CE,ISO & Calibration report are available |
Matched Rotors for TG16E
| Order No | Rotor | Max speed (rpm) | Max Volume(ml) | Max RCF (g) |
| 16E-1 | Angle rotor | 14000 | 4×8PCR | 12070 |
| 16E-2 | Angle Rotor | 16000 | 12×1.5/2ml | 17940 |
| 16E-3 | Angle Rotor | 14000 | 24×1.5/2ml | 17950 |
| 16E-4 | Angle Rotor | 16000 | 10×5ml | 17880 |
| 16E-5 | Angle Rotor | 13000 | 6×10ml | 14190 |
| 16E-6 | Angle Rotor | 12000 | 24 pieces capillary vessel | 15800 |
| 16E-7 | Angle Rotor | 16000 | 40×0.2ml | 19040 |
| 16E-8 | Angle Rotor | 13000 | 40×0.5ml | 17210 |
| 16E-9 | Angle Rotor | 13000 | 30×0.5ml | 13900 |
Features:
1. Brushless motor, no pollution, free-maintenance.
2. Microprocessor control, LCD display which indicates the speed, time, RCF in operation, 10 kinds of brake setting, operate simply.
3. Electric lid lock, super speed, imbalance protection. The centrifuge body is made of high quality steel, stainless steel chamber, safe and reliable.
4. Rotor is connected to spindle by specialized taper sleeve, loading simple and quick, no direction.
5. 3 tiers protection steel cover and get the ideal centrifugation result
HL-dsDNase is especially developed to remove contaminating genomic DNA from RNA preparations. Figure 1 shows that HL-dsDNase can reduce 50 ng of human gDNA to levels non-detectable by qPCR. In figure 2, a human total RNA sample was treated with HL-dsDNase and analysed on the Bio-Rad Experion™ System. The results indicate that HL-dsDNase has minimal impact on RNA quality and quantity.
HL-dsDNase is an engineered version of dsDNase that is rapidly and completely inactivated by incubation for 5 minutes at 58°C with 1 mM DTT at pH 8.0 or above. Chemical inactivation in downstream compatible RT or PCR buffers allows milder heat inactivation and, in some cases, skipping heat inactivation altogether. This makes HL-dsDNase very useful for removal of DNA from RNA preparations since the enzyme may be inactivated using various strategies with reduced risk of auto-degradation of RNA in the presence of magnesium, making HL-dsDNase an ideal enzyme when working with small volumens of RNA.
HL-dsDNase is also an excellent choice for removal of unwanted external DNA from samples prior to analysis of nucleic acids protected by biological membranes, e.g., bacteria, viruses, and sperm. Especially if using downstream analysis methods that might be affected by host cell DNA, as metagenomic sequencing and STR-profiling. The easy inactivation of HL-dsDNase makes it a fast and efficient alternative to methods as differential extraction, where sample material is often lost.
HL-dsDNase is especially developed to remove contaminating genomic DNA from RNA preparations. Figure 1 shows that HL-dsDNase can reduce 50 ng of human gDNA to levels non-detectable by qPCR. In figure 2, a human total RNA sample was treated with HL-dsDNase and analysed on the Bio-Rad Experion™ System. The results indicate that HL-dsDNase has minimal impact on RNA quality and quantity.