Lactobacillus brevis

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

Overview

• Detection kits for H1N1
• CE-IVD marked version available for in vitro diagnostic use
• Available in TaqMan format for analysis

Influenza is caused by three immunologic types of RNA viruses (A, B and C) within the Orthomyxoviridae family. Seasonal influenza is typically caused by three major subtypes of hemaglutinin (H1, H2 and H3) and two subtypes of neuraminidase (N1 and N2). A novel sub-type of influenza A virus called pandemic H1N1 2009 virus was identified in Mexico and reported by the CDC and WHO in April, 2009 (Novel swine-origin influenza A (H1N1) virus investigation team, 2009; CDC, 2009; and Fraser et al., 2009). H1N1 2009 is a novel sub-type virus that transmits easily between humans with 21 countries reporting cases within a month of initial identification (CDC, 2009-b). It is essential that public health laboratories around the world undertake detailed surveillance to monitor the spread and impact of pandemic H1N1 2009 virus as well as try to predict future changes in virulence (Fraser et al., 2009). Methods for the rapid diagnosis, case identification and tracking of this novel pathogen in the human population are therefore required to develop appropriate management strategies to mitigate morbidity and mortality.

H1N1 TaqMan RT-PCR Kit, 100 reactions

• Ready to use format, including Master Mix for the target and PCR control to monitor for PCR inhibition and validate the quality
• Specific Primer and Probe mix for the pathogen/virus/viroid of interest
• Primer and Probe mix
• Positive and negative control to confirm the integrity of the kit reagents

H1N1 TaqMan RT-PCR Probe/Primer Set and Controls, 100 reactions

• Specific Primer/Probe mix and Positive Control for the pathogen/virus/viroid of interest
• Nuclease-free water
• Can be used together with Norgen’s RT-PCR Master Mix (#28113) or customer supplied master mix

Details

Supporting Data

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Storage Conditions and Product Stability
All kit components can be stored for 1 year after the date of production without showing any reduction in performance.

All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.

Component	Cat. TM27950 (100 preps)	Cat. TM27910 (100 rxns)
MDx TaqMan 2X RT-PCR Master Mix	2 x 700 μL	–
H1N1 Primer & Probe Mix	280 μL	280 μL
H1N1 Positive Control	150 μL	3 x 50 μL
Nuclease-Free Water (Negative Control)	1.25 mL	1.25 mL
Product Insert	1	1

Propargyl-PEG6-t-butyl ester is a heterobifunctional linker consisting of an alkyne group and a t-butyl protected carboxyl group. The alkyne group can participate in copper catalyzed azide-alkyne Click Chemistry to form a stable triazole linkage. The t-butyl group can be hydrolyzed under acidic conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Propargyl-PEG6-t-butyl ester is a heterobifunctional linker consisting of an alkyne group and a t-butyl protected carboxyl group. The alkyne group can participate in copper catalyzed azide-alkyne Click Chemistry to form a stable triazole linkage. The t-butyl group can be hydrolyzed under acidic conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.