PEG13-bis(Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane)

Description

Brucella abortus is an intracellular, blood-borne parasite. It is a Gram-negative coccobacillus that causes an infectious and contagious disease called Brucellosis. The disease primarily affects cattle but it can also be transmitted to humans from infected animals and consuming their products. The disease can lead to great economic loss especially in the dairy and agricultural industry. The Brucella abortus genome contains two DNA chromosomes in a circular confirmation; the first chromosome is approximately 2.1 Mb and the second chromosome is approximately 1.2Mb. Unusually it does not contain any plasmids or genomic islands that relate to pathogenicity and lacks many other genes that code for common virulence factors including capsules, fimbriae, exotoxins, cytolysins, resistance forms, or antigenic variation. The most common mode of transmission to humans is through the ingestion of unpasteurized milk and cheese products as the bacteria are present in the milk glands of infected female cows. In cattle transmission can also be through ingestion but in addition, the bacteria can persist in the reproductive tracts of males, namely seminal vesicles, ampullae, testicles, and epididymides, allowing sexual transmission. In humans the bacteria enter macrophages by phagocytosis and then live in compartments of vacuolar space along the endoplasmic reticulum. They persist by inhibiting host apoptosis and go onto form chronic disease causing lesions in the liver, spleen, bone marrow and kidneys. In cattle the bacteria additionally infect the trophoblast epithelial cells, which provide nutrition to the embryo. The trophoblast cells eventually lyse, releasing further bacteria into the blood stream of the embryo. The B. abortus cells in the blood stream go on to colonize the placenta and fetus in pregnant female cows, resulting in abortion of the fetus. Abortion can also result from insufficient anti-Brucella activity in the amniotic fluid. In humans, the disease can be either acute or chronic and some of the symptoms include fluctuating fever, chills, sweating, headache, muscle pain and weight loss. Once a person becomes infected they are prescribed a combination of tetracycline and streptomycin for 3-6 weeks. In cattle, additional symptoms include arthritic joints and retained after-birth.

Exceptional value for money
Rapid detection of all clinically relevant subtypes
Positive copy number standard curve for quantification
Highly specific detection profile
High priming efficiency
Broad dynamic detection range (>6 logs)
Sensitive to < 100 copies of target Accurate controls to confirm findings

Overview

• Convenient ready-to-use solution
• High sensitivity and yield
• Robust amplification
• Available in 3 convenient sizes: 100, 200 and 500 reactions (20 µL vol)
• This product is also a recommended add-on to be used with Norgen’s COVID-19 Primer and Probe Solutions and COVID-19 Positive Control 

Norgen’s 2X One-Step RT-PCR Master Mix is a ready-to-use solution that contains components required for RT-PCR amplification of RNA templates. The mix includes M-MuLV reverse transcriptase, Taq DNA polymerase, dNTPs, reaction buffer, MgCl₂, KCl, and a PCR enhancer/stabilizer. The user needs only to add the template, the primer set and water to the Master Mix to set up the RT-PCR reaction. This convenient 2X One-Step RT-PCR Master Mix reduces the time required to set up PCR reactions and reduces the possibility of contamination, particularly when preparing large numbers of reactions. The optimized master mix allows for robust amplification of RNA templates with high yields of PCR products.

Taq DNA Polymerase is a highly thermostable DNA polymerase that possesses a 5´→ 3´ polymerase activity and a very low 5´→ 3´ exonuclease activity. The source of Taq included with Norgen’s 2X One-Step RT-PCR Master Mix is an E. coli strain with a cloned Taq DNA Polymerase gene from Thermus aquaticus YT-1. M-MuLV Reverse Transcriptase is an RNA-directed DNA polymerase that can synthesize a complementary DNA strand initiating from a primer using either RNA (cDNA synthesis) or single-stranded DNA as a template. The source of the Reverse Transcriptase included with Norgen’s 2X One-Step RT-PCR Master Mix is an E. coli strain with a cloned reverse transcriptase gene from M-MuLV.

Norgen’s 2X One-Step RT-PCR Master Mix is available in 3 convenient sizes:

Cat # 28113 – 100 reactions (sufficient for 100 reactions x 20 µL reaction volume)
Cat # 28114 – 200 reactions (sufficient for 200 reactions x 20 µL reaction volume)
Cat # 28115 – 500 reactions (sufficient for 500 reactions x 20 µL reaction volume)

Details

Supporting Data

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Reagents supplied – Cat # 28115

• 2X One-Step RT-PCR Master Mix (5 Vials, 100 Reactions each) – Sufficient reagent for 500 x 20 µL reactions

Storage Conditions and Product Stability
2X One-Step RT-PCR Master Mix should be stored at -20°C. For everyday use an aliquot can be stored at 4°C for up to 3 months. Repeated freeze-thaw cycles are not recommended. When stored at the proper temperature this reagent is stable for at least 1 year.