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5X Genta Single-tube RT-PCR Master Mix is a ready-to–use mixture for conducting a cDNA synthesis reaction followed by quantitative and/or qualitative PCR with the possibility of detecting results in real time (RT-PCR-RV). 5X Genta Single-tube RT-PCR Master Mix contains all the components necessary for RT-PCR, including Genta REV M reverse transcriptase, Genta TaqF DNA polymerase, dNTP, Mg2+ and reaction buffer. To set up the RT-PCR reaction, only oligonucleotides, matrix and water must be added to the reaction mixture. The chemically inactivated DNA polymerase Genta TaqF allows for preparatory work at room temperature and provides highly specific amplification due to the possibility of a "hot" start. Inhibition of enzyme activity is removed when it is heated at 95 ° C for 15 minutes.

Fusion DNA polymerase is a recombinant polypeptide consisting of a fusion of the thermostable DNA polymerase of Pyrococcus furiosus (Pfu) and the DNA binding protein of the thermophilic archaea species Sulfolobus solfataricus (Sso7d). The Sso7d protein binds to the minor groove of double-stranded DNA and further stabilizes the polymerase-template complex. Due to this, Fusion DNA polymerase has increased processivity, synthesis accuracy, fragment amplification rate, and increased resistance to PCR inhibitors compared to native Pfu DNA polymerase [1]. Fusion DNA polymerase has 5'→3' polymerase activity, 3'→5' exonuclease activity and synthesizes blunt-ended products.
Fusion DNA polymerase is a good choice for routine cloning and can be used to generate long or complex amplicons by PCR.
Examples of using Fusion DNA polymerase are presented at the end of the description.
Fusion DNA polymerase was isolated from an E. coli strain containing a plasmid with a cloned DNA fragment consisting of the fusion genes of the thermostable DNA polymerase of Pyrococcus furiosus (Pfu) and the DNA-binding protein of Sulfolobus solfataricus (Sso7d).
One unit of activity corresponds to the amount of enzyme required to incorporate 10 nmol of dNTP into the acid-insoluble DNA fraction in 30 min at 74°C.

Genta benzonuclease is a nuclease that cleaves all types of both DNA and RNA (single-stranded, double-stranded, linear and annular) to form oligodeoxynucleotides with terminal 5’-monophosphates 3-5 bases long. It is effective in a wide range of working conditions. It has no proteolytic activity. Isolated from a strain of Escherichia coli expressing the nuclease gene from the microorganism Serratia marcescens. Reaction conditions: 20 - 37 °C, optimal buffer: 50 mM Tris-HCl (pH 8.0 at 25 °C); 100 -150 mm NaCl; 1 mM MgCl2.

10-fold buffer for the ligation reaction, compatible with T4 DNA ligase.

A 5-fold reaction buffer that does not contain magnesium sulfate.

Fluorescent probes are an oligonucleotide with a fluorescent label and a suppressor that can bind to the DNA chain during amplification. When the probe is in a free state, both dyes are located close to each other, therefore, fluorescence quenching occurs. During PCR, at the stage of chain elongation, the hybridized probe is cleaved by a polymerase that has 5’-exonuclease activity, the dyes are separated, and a fluorescent signal proportional to the amount of the amplified product is observed. The fluorescence intensity increases in each cycle in proportion to the probe splitting rate. The use of probes in PCR makes it possible to increase the accuracy and selectivity of the reaction. A wide range of fluorescent dyes: FAM, R6G, VIC, HEX, JOE, TAMRA, ROX, Cy 5 and Cy5.5 The possibility of multiplex analysis of multiple DNA targets in one test tube. High efficiency of PCR-RV. Probe quality control: HPLC purification and mass spectrometry verification

The magnetic sorbent ExtraGen® is an aqueous suspension of solid-phase magnetic particles based on iron oxide and silicon oxide. It is designed to isolate nucleic acids from clinical and biological samples. The suspension of the magnetic sorbent has an "intermediate" sedimentation stability, which is especially important when using the material in automatic DNA isolation systems. ExtraGen® particles are resistant to heat, strong oxidants and intense mechanical influences (centrifugation, dispersion, etc.). The use of proprietary buffer solution allows the particles to be stored at room temperature for a year without deterioration of their sorption properties. As tests have shown, the ExtraGen® sorbent shows high efficiency in the isolation of nucleic acids both in manual mode and with the use of automatic stations.

Genta RevM revertase (reverse transcriptase) is a genetically modified reverse transcriptase of the Moloney mouse leukemia virus (MMLV), designed for the synthesis of a complementary DNA chain (cDNA) on a single-stranded RNA matrix. Genta RevM was isolated from a strain of Escherichia coli expressing a modified MMLV reverse transcriptase gene. In comparison with the reverse transcriptase of the "wild type" Genta REV M, the revertase has increased thermal stability: the optimal temperature for the enzyme is 50 ° C, the enzyme remains active at temperatures up to 65 ° C. Reverse transcriptase Genta RevM revertase provides higher cDNA yield, allows longer fragments to be synthesized, has improved efficiency when using GC-rich RNA matrices, and also has higher productivity. The Genta RevM reverse transcriptase is suitable for RT-PCR in the "one-step" format.

The kit is designed for isolating plasmid DNA from bacteria.
The operating principle of the kit is based on alkaline lysis of RNase-treated biomaterial, followed by selective binding of DNA from the clarified lysate on a siliconized column membrane. The colored solutions in the kit allow you to control the correct order of adding components, the completeness of lysis and neutralization.
Benefits of the set:

• Yield up to 25 µg from 2 ml of bacterial culture and up to 60* µg from 6-10 ml of bacterial culture
• Indicators A260/280 = 1.80±0.05, A260/230 ≥ 2.1
• Plasmid DNA has been verified to be suitable for restriction reactions, transformation, sequencing, PCR, transfection.

*product yield depends on the copy number of the plasmid, culture volume and cultivation conditions

Genta UDG thermolabile glycosylase is a thermosensitive enzyme that catalyzes the release of uracil from uracil-containing single- or double-stranded DNA. It is used to prevent the appearance of false positive results caused by contamination with amplicons. The enzyme does not show activity against RNA and oligomers and is fully incubated during the first PCR cycle when heated above 70 ° C, without interfering with the amplification of PCR products.

The kit uses the technology of DNA sorption on magnetic particles. The entire DNA extraction process combines the steps of sample lysis, subsequent selective binding of the released DNA to magnetic beads, and then, after a series of washes, elution with the option of concentrating the DNA in a low-salt buffer.
Benefits of the set:

• High yield of genomic DNA from E.coli and eukaryotic cells: 4-5 µg DNA from 0.5-1*109 bacterial cells and 0.25-1*106 eukaryotic cells.
• High yield of genomic DNA when isolated from various mouse tissues and organs.
• Indicators A260/280 = 1.80±0.05, A260/230 ≥ 2.1
• Purified DNA is verified to be free of proteins, nucleases or other contaminants and can be used in a wide range of molecular biology techniques such as PCR, RT-PCR or other enzyme reactions.
• High throughput for DNA extraction from large numbers of samples.

ExoGen is a ready-to-use mixture of Genta-ExoI exonuclease I enzymes and the thermolabile alkaline phosphatase Genta-ALP in a reaction buffer. ExoGen mixture is used for enzymatic hydrolysis of an excessive amount of primers and nucleotides in PCR products (amplicons). The PCR products purified in this way are suitable for further use in various biological applications, such as cloning and DNA sequencing.