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Home > ÀüÁ¦Ç°º¸±â > Real Time PCR > TB Green °ËÃâ(Clontech) > Terra qPCR Direct TB Green¢â Premix
TB Green¢â

Terra qPCR Direct TB Green¢â Premix

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Clontech
638319
Terra qPCR Direct TB Green¢ç Premix
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200 rxns
462,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x33146, x33145, x99052, x99051

DNA Á¤Á¦ ¾øÀÌ °ð¹Ù·Î Real Time PCR Àû¿ë
Real Time qPCR Master Mix-Terra qPCR Direct TB Green¢â Premix

Terra qPCR Direct TB Green¢â Premix´Â 2X master mix·Î ¸¶¿ì½º ²¿¸®³ª ±Í Á¶Á÷, ¶Ç´Â ½Ä¹°ÀÇ ÀÙ Á¶Á÷À» Á¤Á¦ ¾øÀÌ crude extract ÇüÅ·ΠReal Time PCR (qPCR)¿¡ °ð¹Ù·Î Àû¿ëÇÒ ¼ö ÀÖ´Ù.

qPCR¿¡ »ç¿ëµÇ´Â Terra PCR Direct Polymerase´Â whole cell, crude cell lysate ¶Ç´Â tissue extract±×¸®°í Á¤Á¦µÈ DNA¿¡¼­µµ ¶Ù¾î³­ ÁõÆø È¿À²À» º¸À̵µ·Ï ÃÖÀûÈ­µÈ non-Taq polymerase·Î premix ÇüÅ·ΠÁ¦°ø µÈ´Ù.
±âº»ÀÌ µÇ´Â Terra Polymerase´Â GC ÇÔ·®ÀÌ 70% ÀÌ»óÀÎ À¯ÀüÀÚ Àû¿ë ½Ã¿¡µµ ¶Ù¾î³­ ÁõÆø È¿À²À» ³ªÅ¸³¿À» È®ÀÎ ÇÏ¿´´Ù(Figure 2). º» premix ½Ã¾àÀº TB Green¢â Green I dye »Ó¸¸ ¾Æ´Ï¶ó monoclonal antibody¸¦ »ç¿ëÇÏ¿© hotstart PCRÀ» ÁøÇàÇϱ⠶§¹®¿¡ 10ºÐ°£ÀÇ predeanture time ¾øÀ̵µ automatic hot start ¹ÝÀÀÀÌ °¡´ÉÇÏ´Ù.

Terra qPCR Direct TB Green¢â Premix´Â fluorescence signalÀ» normalizationÇϴµ¥ ÀÌ¿ëµÇ´Â 2Á¾·ùÀÇ ROX reference Dye¸¦ Æ÷ÇÔÇÏ°í ÀÖ´Ù.

ROX Reference Dye LSR
- ROX Reference Dye LSR´Â excitation source°¡ 488 nm laserÀÎ ±â±â¿¡ ÀûÇÕÇÏ´Ù.

ROX Reference Dye LMP
- ROX Reference Dye LMP´Â excitation source°¡ lamp ¶Ç´Â LEDÀÎ ±â±â¿¡ ÀûÇÕÇÏ´Ù.

ROX Reference Dye ÷°¡ ºÒÇÊ¿ä
Takara»çÀÇ Thermal Cycler Dice Real Time System II ¶Ç´Â Cepheid »çÀÇ Smart Cycler System, Roche»çÀÇ LightCycler¿¡´Â º°µµ·Î ÷°¡ÇÒ ÇÊ¿ä°¡ ¾ø´Ù.

±×¸² 1

Figure 1. Real-time PCR with crude extracts-Terra qPCR Direct TB Green¢â 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¢â Premix or a conventional qPCR premix. Using the manufacturer's recommended conditions for each enzyme mix, a 165 bp region of the ¥â-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¢â Premix corresponded to the theoretical quantities of each gene, while the conventional product was clearly affected by inhibitors present in the crude samples.

±×¸² 2

Figure 2. Real-time PCR of GC-rich targets-Terra qPCR Direct TB Green¢â 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¢â Premix (triangles) or one of two conventional TB Green¢â premixes (one specifi¡©cally 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 ¥â-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¢â Premix was the only premix able to consistently amplify all of the targets assayed.
Applications
Perform direct real-time PCR from crude extracts, and dirty samples without having to purify template.
Components
  • 2X Terra qPCR Direct TB Green¢â Premix* : Contains Terra PCR Direct Polymerase (w/ hot start Ab), dNTP Mixture, Mg2+ and TB Green¢â Green I dye.
  • ROX Reference Dye LSR (50X)
  • ROX Reference Dye LMP (50X)
Storage Conditions
Store all components at -80¡É after thawing, store the kit-protected from light at 4¡É and use within 6 months.
Do not refreeze!

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