Terra qPCR Direct TB Green Premix
Terra qPCR Direct TB Green Premix
Brand: Takara Bio.
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Terra qPCR Direct TB Green Premix
Terra qPCR Direct TB Green Premix
Terra qPCR Direct TB Green Premix is a 2X master mix designed specifically for real-time PCR with TB Green chemistry. In addition to TB Green dye, this mix contains Terra qPCR Direct Polymerase, a novel enzyme developed for optimal amplification from crude or dirty templates; it’s perfect for amplifying short DNA targets (up to 2 kb), regardless of GC content or template purity. The premix also contains a monoclonal antibody that suppresses polymerase activity up to 98°C, allowing automatic hot start PCR.
Overview
- Perform real-time PCR with crude tissue extracts or dirty samples—no template purification necessary
- Easily amplify GC-rich targets
- Convenient 2X master mix contains TB Green dye
- Optimized for use with the laser-, lamp-, or LED-based real-time instrument of your choice
- Automatic hot start
- Perfect for high-throughput screening applications
Applications
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Perform direct real-time PCR from crude extracts and dirty samples, without having to purify your template
Real-time PCR with crude extracts—Terra qPCR Direct TB Green Premix versus a conventional 2X qPCR premix
Real-time PCR with crude extracts—Terra qPCR Direct TB Green Premix versus a conventional 2X qPCR premix. Real-time PCR was performed using undiluted, 4X diluted, and 16X diluted crude alkaline-heat extracts of mouse spleen or cow muscle (beef foodstuff), and either Terra qPCR Direct TB Green Premix or a conventional qPCR premix. Using the manufacturer's recommended conditions for each enzyme mix, a 165 bp region of the beta-globin gene Hbb-b1 was amplified from the mouse spleen extract (Panel A), and a 289 bp region of the cytochrome c oxidase gene (COX1) was amplified from the beef extract ( Panel B). Data generated by Terra qPCR Direct TB Green Premix corresponded to the theoretical quantities of each gene, while the conventional product was clearly affected by inhibitors present in the crude samples.
Real-time PCR of GC-rich targets—Terra qPCR Direct TB Green Premix versus conventional 2X qPCR premixes
Real-time PCR of GC-rich targets—Terra qPCR Direct TB Green Premix versus conventional 2X qPCR premixes. Targets with GC-content greater than 70% were amplified by real-time PCR using either human testis cDNA (equivalent to 50 ng–5 pg of total RNA; Panel A) or human genomic DNA (100 ng–10 pg; Panel B) as template, and either Terra qPCR Direct TB Green Premix (triangles) or one of two conventional SYBR premixes (one specifically for GC-rich targets; see figure legend). Using the manufacturer's recommended conditions for each enzyme mix, gene-specific primers were used to amplify portions of the jun-D proto-oncogene (JUND), the BTB domain-containing protein 6 gene (BTBD6), and the cyclin I gene (CCNI) from the cDNA (Panel A); and a portion of the beta-actin CpG island (ACTB_CpG) from the genomic DNA (Panel B). The resulting Ct values were plotted against the initial quantity of DNA used in each assay. Terra qPCR Direct TB Green Premix was the only premix able to consistently amplify all of the targets assayed.
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Real-time PCR with crude extracts—Terra qPCR Direct TB Green Premix versus a conventional 2X qPCR premix
Real-time PCR with crude extracts—Terra qPCR Direct TB Green Premix versus a conventional 2X qPCR premix. Real-time PCR was performed using undiluted, 4X diluted, and 16X diluted crude alkaline-heat extracts of mouse spleen or cow muscle (beef foodstuff), and either Terra qPCR Direct TB Green Premix or a conventional qPCR premix. Using the manufacturer's recommended conditions for each enzyme mix, a 165 bp region of the beta-globin gene Hbb-b1 was amplified from the mouse spleen extract (Panel A), and a 289 bp region of the cytochrome c oxidase gene (COX1) was amplified from the beef extract ( Panel B). Data generated by Terra qPCR Direct TB Green Premix corresponded to the theoretical quantities of each gene, while the conventional product was clearly affected by inhibitors present in the crude samples.
Real-time PCR of GC-rich targets—Terra qPCR Direct TB Green Premix versus conventional 2X qPCR premixes
Real-time PCR of GC-rich targets—Terra qPCR Direct TB Green Premix versus conventional 2X qPCR premixes. Targets with GC-content greater than 70% were amplified by real-time PCR using either human testis cDNA (equivalent to 50 ng–5 pg of total RNA; Panel A) or human genomic DNA (100 ng–10 pg; Panel B) as template, and either Terra qPCR Direct TB Green Premix (triangles) or one of two conventional SYBR premixes (one specifically for GC-rich targets; see figure legend). Using the manufacturer's recommended conditions for each enzyme mix, gene-specific primers were used to amplify portions of the jun-D proto-oncogene (JUND), the BTB domain-containing protein 6 gene (BTBD6), and the cyclin I gene (CCNI) from the cDNA (Panel A); and a portion of the beta-actin CpG island (ACTB_CpG) from the genomic DNA (Panel B). The resulting Ct values were plotted against the initial quantity of DNA used in each assay. Terra qPCR Direct TB Green Premix was the only premix able to consistently amplify all of the targets assayed.
