Escherichia coli (E.coli) O157:H7 Quantified Bacterial DNA Standards
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Detail
Overview
Quantified standard to be used as a positive control or PCR quantification standard
Vigorously quantified using multiple methods
Includes partial sequence of STX-1 subunit A, and STX-2 subunit A genes (please refer to product insert for more details)
Compatible with E. coli O157:H7 TaqMan PCR Detection Kit and E. coli O157:H7 TaqMan PCR Detection Probe/Primer and Control Set
E. coli O157:H7 is a rod-shaped, gram negative bacterium. It is an enterohemorrhagic strain of the common E. coli bacterium and infection by the O157:H7 strain is commonly associated with hemorrhagic colitis. E. coli O157:H7 is recognized by its somatic (cell wall) antigen (O157) and its flagella antigen (H7). In addition, E. coli O157:H7 is known to produce Shiga-like toxins, which cause severe symptoms. While most patients can recover from the infection, up to 15% of the patients may develop hemolytic uremic syndrome, a type of kidney failure that could be fatal. Infection of E. coli O157:H7 usually results from consumption of poorly prepared food including undercooked meat (particularly ground beef), untreated water or raw unpasteurized milk.
Norgen’s E. coli O157:H7 Quantified Bacterial DNA Standards are cloned fragments of the two Shiga-like toxin regions purified using Norgen’s sample preparation technology. They include partial sequences of STX-1 subunit A gene and STX-2 subunit A gene. Please refer to Appendix A for sequence information. The purified DNA is quantified vigorously using multiple methods including spectrophotometry, gel densitometry and real-time PCR. It is intended to be used as positive controls or PCR quantification standards for E. coli O157:H7. This product is compatible with Norgen Biotek’s E. coli O157:H7 Taqman PCR Detection Kit (Cat. TM41350) and E. coli O157:H7 Taqman PCR Detection Probe/Primer and Control Set (Cat. TM41310).
Storage Conditions Upon receipt, store Norgen’s E. coli O157:H7 Quantified Bacterial DNA Standards at -20°C or lower. Avoid multiple freeze-thaw cycles. If needed, prepare smaller working aliquots and store at -20°C or lower.
Sal-Like Protein 4 (SALL4), is a zinc finger transcription factor found in germ cells and human blood progenitor cells, with functional involvement in modulating Oct-4 to maintain embryonic stem cell pluripotency. SALL4 is a useful marker for acute myeloid leukemia, B-cell acute lymphocytic leukemia, intratubular germ cell neoplasia, seminomas/dysgerminomas, and yolk sac tumors (both pediatric and postpubertal). Anti-SALL4 is used to detect embryonal carcinomas, hepatocellular carcinoma (HCC), gliomas, ovarian primitive germ-cell tumors, choriocarcinomas, spermatogonia, teratoma, gastric cancer, breast cancer, and lung cancer. Expression of SALL4 is often associated with poor prognosis in HCC, and with metastasis in endometrial cancer, colorectal carcinoma, and esophageal squamous cell carcinoma.
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
