[QP5520] Q-PAGE™ TGN Precast Gel (Midi, 15 wells, 4-15%), 10 gels

Description 

The RNAok™ RNase Inhibitor is a recombinant mammalian RNase inhibitor which possesses very high affinity for eukaryotic pancreatic-type ribonuclease. The RNAok™ RNase Inhibitor forms a 1:1 complex with pancreatic RNase A by noncovalent binding, presenting a noncompetitive inhibitory activity on these pancreatic enzymes. RNAok™ RNase Inhibitor is active against RNase A, RNase B, RNase C but not RNAse H, RNase I, RNase T1, RNase T2, and S1 nuclease. RNAok™ RNase Inhibitor is compatible with RT-PCR enzymes such as AMV, M-MLV and ExcelRT™ Reverse Transcriptase or Taq DNA polymerase.

Application

•  cDNA Synthesis
• in vitro translation
• in vitro transcription
• One-step RT-PCR
• Separation and identification of specific ribonuclease activities

Storage Buffer

40 mM HEPES-KOH (pH 7.5), 100 mM KCl, 8 mM DTT,   0.1 mM EDTA, stabilizer and 50% (v/v) glycerol

Storage

-20°C for 24 months 

The RNAok™ RNase Inhibitor is a recombinant mammalian RNase inhibitor which possesses very high affinity for eukaryotic pancreatic-type ribonuclease. The RNAok™ RNase Inhibitor forms a 1:1 complex with pancreatic RNase A by noncovalent binding, presenting a noncompetitive inhibitory activity on these pancreatic enzymes. RNAok™ RNase Inhibitor is active against RNase A, RNase B, RNase C but not RNAse H, RNase I, RNase T1, RNase T2, and S1 nuclease. RNAok™ RNase Inhibitor is compatible with RT-PCR enzymes such as AMV, M-MLV and ExcelRT™ Reverse Transcriptase or Taq DNA polymerase.

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 5 x 109 AAV particles as determined by qPCR
AAV Vector Serotype	Any
Average Recovery	> 80%
Input Type	Cells, media, or mixed
Input Volume	0.5 mL – 8 mL
Minimum Elution Volume	200 µL
Time to Complete Purifications	1 – 2 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