Neisseria gonorrhoeae Quantified Bacterial DNA Standard
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Detail
Overview
Quantified standard to be used as a positive control or PCR quantification standard
Vigorously quantified using multiple methods
Neisseria gonorrhoeae is a Gram-negative coccus of the Neisseria genus. N. gonorrhoeae is usually seen in pairs infecting human cells. It has a circular DNA genome of approximately 1Mbp encoding over 2000 genes. N. gonorrhoeae is the etiological agent of the sexually transmitted infection (STI) gonorrhoea, which globally causes an estimated 60 million new cases of gonococcal disease annually. It is second only to Chlamydia trachomatis as the most reported notifiable sexually transmitted disease. Infections with N. gonorrhoeae are primarily restricted to the mucus membranes of the endocervix, urethra, rectum, and pharynx. In females, gonorrhoea is a major cause of pelvic inflammatory disease and may lead to tubal infertility, ectopic pregnancy, and chronic pelvic pain, whereas in males it primarily causes urethritis. Importantly, these infections may often be asymptomatic, thereby contributing to further transmission and maintenance of the disease within populations.
Norgen’s Neisseria gonorrhoeae Quantified Bacterial DNA Standard is prepared from pelleted bacteria grown on culture plates using Norgen’s sample preparation technology. The purified DNA is quantified vigorously using multiple methods including spectrophotometry, gel densitometry and real-time PCR. It is intended to be used as a positive control or PCR quantification standard for Neisseria gonorrhoeae.
Volume Provided – 250 µL DNA Quantity – 2 x 104 copies per µL
Storage Conditions Upon receipt, store Norgen’s Neisseria gonorrhoeae Quantified Bacterial DNA Standard at -20oC or lower. Avoid multiple freeze-thaw cycles. If needed, prepare smaller working aliquots and store at -20oC or lower.
Other Products
HCM131 Sorbitol MacConkey Agar Base
Product Info
Document
Product Info
Introduction
Intended Use
For the isolation and differentiation of sorbitol-negative Escherichia coli serotype O157 from food and animal feed and other materials
Principle and Interpretation
Enzymatic digest of casein and enzymatic digest of animal tissue provides nitrogen source, vitamins and growth factors; sodium chloride maintains balanced osmoticpressure; sorbitol is a fermentable sugar; No. 3 bile salt and crystal violet inhibit Gram-positive bacteria, potassiumtellurite inhibits non-o157 Escherichia coli, and cefixime inhibits Proteus; neutral red is a pH indicator; and agar is the coagulant of the culture medium.
Formulation
Ingredients
/liter
Enzymatic digest of casein
17.0g
Enzymatic digest of animal tissue
3.0g
Sorbitol
10.0g
Bile salt No.3
1.5g
Sodium chloride
5.0g
Neutral red
0.03g
Crystal violet
0.001g
Agar
15.0g
pH 7.1±0.2 at 25°C
Preparation
Weigh 51.5g of this product, add 1L of distilled water or deionized water, stir, heat and boil until completely dissolved, dispense into Erlenmeyer flasks, and sterilize at 121℃ for 15min. Melt the sterilized culture medium and cool it to about 45℃, add 1 tube of supporting reagent (SR0240) (0.25mg potassium tellurite and 0.005mg Cefixime) to every 100mL of basal culture medium, mix and pour into the plate.
Quality Control
Cultural characteristics observed after incubation at 35-37°C for 18-24hours
Quality control strains
Growth
Colony color
Escherichia coli ATCC25922
PR≥0.7
pale pink to red
Escherichia coli 0157:H7 NCTC12900
PR≥0.7
colorless
Staphylococcus aureus ATCC 25923
Total inhibition
–
Sorage and Shelf Life
Keep container tightly closed, store in a cool, dry place, away from bright light. Storage period of 3 years.
Precautions
1. When weighing the dehydrated medium, please wear masks to avoid causing respiratory system discomfort
2. Keep container tightly closed after using to prevent clumping.
Waste Disposal
Microbiological contamination was disposed by autoclaving at 121°C for 30 minutes.
Document
Intended Use For the isolation and differentiation of sorbitol-negative Escherichia coli serotype O157 from food and animal feed and other materials Principle and Interpretation Enzymatic ……
High yield and high purity DNA ready for any application
Available in a variety of formats to properly suit your needs
Compatible with blood collected on a variety of commercially available tubes
These kits allow for the isolation of DNA from the blood of various species, including humans and will recover genomic DNA, mitochondrial DNA, viral DNA and bacterial DNA. The purified DNA is of excellent quality and yield and completely compatible with any downstream application including PCR, qPCR and genotyping.
Blood DNA Isolation Mini Kit
Norgen’s Blood DNA Isolation Mini Kit is designed for the rapid preparation of DNA from up to 200 µL of whole blood using a rapid spin column protocol. Preparation time for a single sample is less than 30 minutes and each kit contains sufficient materials for 50 preparations.
Blood DNA Isolation Midi Kit
Norgen’s Blood DNA Isolation Kit is designed for the rapid spin column preparation of DNA from 0.3 to 2 mL volumes of whole blood. Preparation time for a single sample is less than 30 minutes, and each kit contains sufficient materials for 20 preparations.
Blood DNA Isolation Maxi Kit
This kit is designed for the rapid preparation of DNA from 3 mL up to 10 mL of whole blood. Preparation time for a single sample is less than 30 minutes.
Blood DNA Isolation 96-Well Kit (High Throughput)
This kit provides a rapid method for the high-throughput isolation of DNA from up to 200 µL of whole blood. Fast and easy processing using either a vacuum manifold or centrifugation.
Blood DNA Isolation Kit (Magnetic Bead System)
Norgen’s Blood DNA Isolation Kit (Magnetic Bead System) is designed for the rapid preparation of genomic DNA from up to 200 µL of whole blood from various species, including human. Purification is based on magnetic beads as the separation matrix. Norgen’s magnetic beads bind DNA under optimized salt concentrations and releases the bound DNA under low salt and slightly alkali conditions. The genomic DNA is preferentially purified from other cellular proteinaceous components. Typical yields of genomic DNA will vary depending on the cell density of the blood sample. The purified genomic DNA is fully digestible with all restriction enzymes tested, and is completely compatible with downstream applications including real-time PCR, NGS and microarray analysis.
Blood DNA Isolation 96-Well Kit (High Throughput Magnetic Bead System)
96-well format for high throughput applications. Purification with the 96-well plates can be integrated with a robotic automation system.
40 minutes (hands-on time Cat. 59800) 50 minutes (hands-on time Cat. 62600)
* Average DNA yield will vary depending on the donor
Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment. The kit contains a ready-to-use Proteinase K, which is dissolved in a specially prepared storage buffer. The buffered Proteinase K is stable for up to 1 year after the date of shipment when stored at room temperature.
Double-Strand Specific dsDNase (dsDNase) is ideal for fast and effective removal of contaminating DNA from PCR master mixes.
Taq polymerases are commonly contaminated by bacterial DNA. This is a problem in PCR based bacterial typing and detection as it might cause false positive results. The unique properties of dsDNase make it suited for removal of contaminating DNA from PCR master mixes prior to addition of DNA template.
In figure 1, a PCR master mix was treated with different amounts of dsDNase before performing a qPCR to measure the contaminating bacterial DNA in the master mix. ArcticZymes dsDNase effectively removed contaminating DNA below known levels of the assay detection limits.
The dsDNase from Arctic shrimp (Pandalus borealis) is recombinantly produced in Pichia pastoris. It cleaves phosphodiester linkages in DNA to yield oligonucleotides with 5’-phosphate and 3’-hydroxyl termini.
The specific activity is estimated to be 30 times higher than that of bovine DNase I. In the presence of magnesium as only divalent cation and using oligos as a substrate, the activity towards dsDNA is 5000-fold higher than towards ssDNA.
The unique double strand-specificity allows specific degradation of dsDNA while leaving shorter ssDNA as primers and probes essentially intact. Easy inactivation by moderate heat (65°C) allows addition of DNA intended for analysis directly after removal of contaminating DNA.
Can be heat-inactivated by moderate heat treatment (65°C for 15 minutes)
Producing 5′-phospho-oligonucleotide products
Figures
Figure 1. The dsDNase effectively removes contaminated DNA
The dsDNase effectively removes contaminated DNA:
A PCR master mix was preincubated with various concentrations of dsDNase. After treatment, no DNA was amplified in non-template controls.
Properties
Specificity towards double-stranded DNA
Nucleic acid specificity has been tested towards double- and single-stranded DNA and RNA oligonucleotides. The specificity of dsDNase towards the substrate has been measured using 15-mer oligonucleotides with FAM at 5′ and DarkQuencher® 3′ (Eurogentec). The fluorescence is proportional to enzyme activity. Assay conditions: 25 mM Tris pH 7.5, 5 mM MgCl2, and 2 μM oligonucleotide.
Double-Strand Specific dsDNase (dsDNase) is ideal for fast and effective removal of contaminating DNA from PCR master mixes.
Taq polymerases are commonly contaminated by bacterial DNA. This is a problem in PCR based bacterial typing and detection as it might cause false positive results. The unique properties of dsDNase make it suited for removal of contaminating DNA from PCR master mixes prior to addition of DNA template.
