Genomic DNA can be isolated from as few as 10 bacterial cells in 1 mL of milk
Isolate genomic DNA from both Gram-negative and Gram-positive bacteria in milk
Can process challenging samples such as mastitic milk
Inhibitor-free DNA is ready for PCR, qPCR, Southern Blot, sequencing & more
Fast and efficient spin-column format
This kit provides a rapid spin column method for the isolation and purification of genomic DNA from both Gram-negative and Gram-positive bacteria in milk samples. It can also be used to process challenging milk samples such as Mastitic milk for example. The kit is highly sensitive and can isolate DNA from a cell density of as little as 10 bacteria contained in 1 mL of milk. Genomic DNA is isolated in 45 minutes, free of inhibitors and ready for any number of downstream applications including PCR, qPCR and Southern Blot analysis, sequencing and more.
*The range of the DNA yield will vary depending upon a number of factors including bacterial species and type of milk (fat content and %)
Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. The Lysozyme should be stored at -20°C upon arrival, and the Resuspension Solution A should be stored at -20°C after addition of the lysozyme. The lyophilized Proteinase K should be stored at -20°C upon arrival and after reconstitution. This kit is stable for 1 year from the date of shipment.
Component
Cat. 21550 (50 preps)
Resuspension Solution A
6 mL
Buffer SK
60 mL
Wash Solution A
18 mL
Elution Buffer B
15 mL
Proteinase K
12 mg
Lysozyme (powder)
120 mg
Spin Columns
50
Collection Tubes
50
Elution Tubes (1.7 mL)
50
Product Insert
1
Other Products
AAV Purification Kits
Product Info
Document
Product Info
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.
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
PACE (PCR Allelic Competitive Extension) genotyping chemistry is a homogeneous, PCR-based allele-specific technology for the analysis of DNA sequence variants, most commonly SNPs (Single Nucleotide Polymorphisms) and Indels (insertion / deletions).
PACE genotyping chemistry is comprised of two parts:
PACE Genotyping Assay: two allele-specific forward primers and one common, reverse primer. The allele-specific forward primers each have different 3′ terminal bases reflecting the target variant, and a unique 5’ tail sequence which is incorporated as part of the fluorescent signal mechanism.
PACE Genotyping Master Mix: containing all remaining components required for PCR and the generation of fluorescent signals. PACE Genotyping Master Mix contains a novel, universal, fluorescent reporting cassette to produce machine-readable fluorescent signals (FAM and HEX) corresponding to the assay genotypes.
When combined with sample DNA, these components create a PACE Genotyping Reaction, as illustrated in the figure below.
We have extensive knowledge and experience in assay design, especially when it comes to allele-specific PCR. PACE Genotyping Assays are available to purchase either Validated and Unvalidated. Validated assays require customer DNA to validate and optimise, for guaranteed performance. Unvalidated assays are designed in silico and supplied untested.
REQUIRED COMPONENTS
qPCR machine or Thermocycler + Fluorescent plate reader
PCR plate or equivalent and appropriate optically clear seal
Template DNA
PCR-grade water
PACE Genotyping Master Mix or PACE 2.0 Genotyping Master Mix
STEPS TO YOUR PACE GENOTYPING ASSAY DESIGN
Place your order on this page. Our support team will contact you by email.
Fill out an Assay Design Template form with all the information we need to process your custom PACE Genotyping Assay order. We will email you a copy of the template when we first contact you, or your can download a copy here.
Using the information you provide us, we will create your PACE Genotyping Assay designs, order the oligos, and send you design sequences.
Once we receive the oligos, we assemble the assay(s) and then ship an aliquot to you (unvalidated) or test on your DNA samples before shipping the aliquot to you (validated).
