Master Mix and Assays

tinto rang is a minor groove / external binding nucleic acid dye that has a high affinity and rapid binding to DNA. It can find potential applications in the making of safe, fluorescence-based Minor Groove Binder (MGB) dyes and probes that have several advantages over DNA intercalating probes in terms of increased stability, sensitivity and specificity. Further leading to make safer and efficient master mixes.

tinto rang™ nucleic acid stain (DNA/RNA) with additional optimization of reaction conditions and primer design, can be used for downstream applications in wide selection of high-quality PCR master mixes, dyes and probes.

High specificity and increased yield shows no reduction in sensitivity upon boiling with agarose and is applicable over a wide pH range, which proves its stability; versatile formulation for broad range of amplicons; convenient room-temperature reaction set-up. Refrigerating at 4° is necessary only during long term storage; dye with agarose gel, in sachets, optimized versions with buffers enable direct gel loading; Super fast: 1-5 minutes reaction time; Super sensitivite; resistance into a variety of reaction inhibitors.

PCR (polymerase chain reaction) is an indispensable tool for molecular biology research. It is used daily in laboratories around the world in a wide array of applications such as cloning, gene expression analysis, genotyping, sequencing, and mutagenesis. tinto rang™ is compatible with common PCR additives such as DMSO and betaine. These additives neither change the color nor affect dye migration. We have examined the gel result stanied with tinto rang™ of PCR product (560bp) amplified using Applied Biosystems 2720 Thermal Cycler using GE ImageQuant LAS-4000 GelDoc System.
Reverse transcription is the synthesis of single-stranded DNA (complementary DNA, or cDNA) using single-stranded RNA as a template, mediated by reverse transcriptases (RTs). The cDNA can be used as a template for amplification by PCR or to generate a cDNA library. Choosing the correct reverse transcriptase for cDNA synthesis is critical to obtaining high yields of quality, full-length cDNA with full gene representation. The ideal RT will reverse-transcribe even the most difficult types of RNA, such as RNA samples from plants (which are normally degraded, resulting in fewer copies of RNA transcripts). Visualisation of the RT-PCR product stained with tinto rang™ after amplification using Applied Biosystems 2720 Thermal Cycler has been observed under GE ImageQuant GelDoc System.
A Real-Time polymerase chain reaction(Real-Time PCR) is a laboratory technique of molecular biology based on the polymerase chain reaction (PCR), it monitors the amplification of a targeted DNA molecule during the PCR. In conventional PCR, the amplicon, is detected in an end-point analysis. In real-time PCR, the accumulation of amplification product is measured as the reaction progresses, in real time, with product quantification after each cycle. The measured fluorescence is proportional to the total amount of amplicon; the change in fluorescence over time is used to calculate the amount of amplicon produced in each cycle.

Applications of Real-Time PCR/qPCR Assays:

Real-time PCR/qPCR assays have become the tool of choice for the rapid and sensitive determination and quantitation of nucleic acid in various biological samples, with diverse applications such as gene expression analysis, the detection of genetically modified organisms in food, and cancer phenotyping.

In research laboratories, qPCR assays are widely used for the quantitative measurement of gene copy number (gene dosage) in transformed cell lines or the presence of mutant genes.

In combination with reverse-transcription PCR (RT-PCR), qPCR assays can be used to precisely quantitate changes in gene expression, for example, an increase or decrease in expression in response to different environmental conditions or drug treatment, by measuring changes in cellular mRNA levels.

Various assays, master mixes and kits using tinto rang dye and probes can be customized for specific applications and techniques.