Introduction

MagZol Reagent is a reagent system for the isolation of total RNA from cells and tissues. The reagent, a single-phase solution consisting of phenol and guanidine isothiocyanate, is modification of the single-step RNA isolation method developed by Chomczynski and Sacchi. The sample is homogenized and lysed in MagZol Reagent which maintains the integrity of the RNA, while disrupting and denaturing endogenous RNases and other cellular components. Extraction of the lysate with chloroform further denatures proteins and separates the mixture into an organic and an aqueous phase. RNA remains exclusively in the aqueous phase, and is subsequently recovered by isopropanol.

This method is suitable for small quantities of tissue (<100mg) and cells (<5 X10⁶), and large quantities of tissue (up to 1g) and cells (<10⁸), of human, animal, plant, or bacterial origin. The simplicity of the MagZol Reagent method allows simultaneous processing of a large number of samples. The entire procedure can be completed in one hour. Total RNA prepared in this manner can be used for Northern blot analysis, dot blot hybridization, poly(A) + selection, in vitro translation, RNase protection assay, and molecular cloning. For use in amplification by thermal cycling, treatment of the isolated RNA with RNase-free DNase I is recommended when the two amplimers lie within a single exon.

Details

Specifications

Advantages

• Flexible – sample amount can be adjusted according to the demand
• Cost performance -the most economical nucleic acid extraction technology

Storage and Stability

MagZol Reagent should be stored at 2-8°C upon arrival and is stable for at least 24 months under the condition. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect its performance.

MagZol Reagent is a reagent system for the isolation of total RNA from cells and tissues. The reagent, a single-phase solution consisting of phenol and guanidine isothiocyanate, is modification of the single-step RNA isolation method developed by Chomczynski and Sacchi. The sample is homogenized and lysed in MagZol Reagent which maintains the integrity of the RNA, while disrupting and denaturing endogenous RNases and other cellular components. Extraction of the lysate with chloroform further denatures proteins and separates the mixture into an organic and an aqueous phase. RNA remains exclusively in the aqueous phase, and is subsequently recovered by isopropanol.

OverView

ArcticZymes RNA to DNA Ligase (ArcticZymes R2D Ligase™) is the first ligase on the market that is able to ligate DNA to 5’-phosphorylated ends of RNA in the presence of a DNA template positioning the joinable ends. With its unique substrate specificity, ArcticZymes R2D Ligase allows the development of new technologies in molecular biology research, diagnostics, and manufacturing.

Key Features:

• Novel substrate specificity
• RNA to DNA ligation
• High purity
• Detergent free
• Efficient ligation of DNA to 5’-phosphorylated ends of RNA in the presence of a DNA template positioning the ligatable ends of DNA and RNA
• ATP dependent dsDNA ligase
• Utilizes Mg²⁺ or Mn²⁺

Suggested Applications:

• RNA capturing
• In vitro transcription  
• NGS library prep
• Transcriptome amplification
• miRNA detection/analysis
• RNA splint ligation

ArcticZymes RNA to DNA Ligase (ArcticZymes R2D Ligase™) is the first ligase on the market that is able to ligate DNA to 5’-phosphorylated ends of RNA in the presence of a DNA template positioning the joinable ends. With its unique substrate specificity, ArcticZymes R2D Ligase allows the development of new technologies in molecular biology research, diagnostics, and manufacturing.

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