Escherichia coli (E.coli) O157:H7 Quantified Bacterial DNA Standards

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).

Details

Supporting Data

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E. coli O157:H7 Quantified Bacterial DNA Standards Figure 1

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Volume Provided	250 μL (for STX1)250 μL (for STX2)
DNA Quantity	2 x 10⁴ copies per μL

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.

Features	Specifications
Main Functions	Isolation total RNA from tissue, cell
Applications	RT-PCR, cDNA synthesis, second generation sequencing
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Adaptive instrument	Nucleic acid extractor, pipetting workstation
Sample type	Tissues, cells, lymphocytes and other clinical sample
Sample amount	Cells grown in suspension:3~5 x 10⁶Animal tissue: 10~20mgPlant tissue: ≤100mg

Contents	IVD3020
Purification Times	200 Preps
MagPure RNA Particles	7 ml
DNase I	4 x 600 µl
DNase Buffer	80 ml
RTL Lysis Buffer	150 ml
Buffer MCB*	75 ml
Buffer MW1*	110 ml
Buffer MW2*	50 ml
RNase Free Water	60 ml

Cat.No	Reagent	IVD3020-F-96
DNase Buffer		60 ml
DNase I		2 x 600 μl
RTL Lysis Buffer		80 ml
Buffer MCB		18 ml
96-Tip		1
Sample plate (DW Plate)	500μl Buffer MCB	1
Wash 1 Plate (DW Plate)	700μl Buffer MW130μl MagPure RNA Partilces	1
DNase Plate	Empty
Wash 2 Plate (DW Plate)	700μl Buffer MW1	1
Wash 3 Plate (DW Plate)	900μl Buffer MW2	1
Elution plate (DW Plate)	80μl RNase Free Water	1

Cat.No	Reagent	IVD3020-TL-06
Purification times		96 Preps
DNase I		2 x 600 μl
DNase Buffer		60 ml
RTL Lysis Buffer		60 ml
Buffer MCB		40 ml
96-Tip		12 PCS
2.0ml V-bottom plate	Row 1/7：500μl Buffer MCBRow 2/8：500μl Buffer MW1Row 3/9：emptyRow 4/10：30μl Magpure RNA Particles500μl Buffer MW2 Row 5/11：900μl Buffer MW2 Row 6/12：80μl RNase Free Water	6

Kit Specifications
Column Binding Capacity	25 µg
Size of Plasmids Purified	Up to 13,000 bp
Average Yield from 1.5 mL of Culture	Up to 20 µg
Time to Complete 10 Purifications	30 minutes

Escherichia coli (E.coli) O157:H7 Quantified Bacterial DNA Standards

Detail

Overview

Details

Supporting Data

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Component	Cat. 13300 (50 preps)	Cat. 46400 (250 preps)	Cat. 46500 (4 preps)	Cat. 46600 (20 preps)	Cat. 60300 (50 preps)	Cat. 63000 (192 preps)
Resuspension Solution AZ	12 mL	60 mL	20 mL	100 mL	12 mL	2 x 20 mL
Lysis Buffer N	40 mL	80 mL	40 mL	2 x 80 mL	40 mL	2 x 40 mL
Buffer TN	20 mL	130 mL	55 mL	2 x 130 mL	20 mL	1 x 55 mL 1 x 20 mL
Wash Solution E	12 mL	2 x 18 mL	–	–	–	–
Elution Buffer K	8 mL	30 mL	–	–	8 mL	2 x 8 mL
Wash Solution J	–	–	25 mL	3 x 25 mL	–	–
Elution Buffer J	–	–	24 mL	120 mL	–	–
RNase A	1 vial	1 vial	1 vial	1 vial	1 vial	1 vial
Magnetic Bead Suspension	–	–	–	–	1 x 1.1 mL	4 x 1.1 mL
Spin Columns	50	250	–	–	–	–
Collection Tubes	50	250	–	–	–	–
DNA Maxi Spin Columns with Collection Tubes (Clear ring in column)	–	–	4	20	–	–
Maxi Spin Filter Columns with Collection Tubes (Grey ring in column)	–	–	4	20	–	–
96-Well Plate	–	–	–	–	–	2
Elution Tubes (1.7 mL)	50	250	–	–	50	–
Elution Tubes (50 mL)	–	–	4	20
96-Well Elution Plate	–	–	–	–	–	2
Adhesive Tape	–	–	–	–	–	2
Product Insert	1	1	1	1	1	1

Kit Specifications
Sample Volume Range	250 µL
Size of RNA purified	All sizes, including small RNA (<200 nt)
Minimum Elution Volume	75 µL
Maximum Elution Volume	100 µL
Time to complete 96 purifications	30 minutes
Average Yield	≥ 1 µg **varies from sample to sample

Component	Cat. Dx69100 (50 preps)	Cat. Dx69300 (192 preps)
Lysis Buffer A	30 mL	100 mL
Solution WN	18 mL	55 mL
Wash Solution A	18 mL	2 x 38 mL
Elution Solution A	6 mL	20 mL
Mini Spin Columns	50	–
96-Well Isolation Plate (Deep Well)	–	2
Collection Tubes	50	–
96-Well Collection Plate (Deep Well)	–	2
Elution Tubes (1.7 mL)	50	–
96-Well Elution Plate (Deep Well)	–	2
Adhesive Tape	–	4
Product Insert	1	1