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Home > ÀüÁ¦Ç°º¸±â > Real Time PCR > Overview > Dye-based Real Time PCR Premix

Dye-based Real Time PCR Premix

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Dye-based (TB Green¢ç) Real Time PCR Premix, ½ÇÇè¿¡ µû¶ó ¼±ÅÃÇϼ¼¿ä!
Introduction
Real Time PCR (Quantitative PCR; qPCR)Àº À¯ÀüÀÚ¹ßÇöÀÇ Á¤±³ÇÑ ºÐ¼®À» À§ÇØ ÀϹÝÀûÀ¸·Î ³Î¸® »ç¿ëµÇ´Â ºÐÀÚ»ý¹°ÇÐÀû ±â¹ýÀÌ´Ù. ´ÙÄ«¶ó¿¡¼­ ÆǸÅÇÏ°í ÀÖ´Â Dye-based Real Time PCR kit´Â ´ÙÄ«¶óÀÇ µ¶ÀÚÀûÀÎ TB Green¢ç intercalating dye¸¦ »ç¿ëÇÏ¿© °í°¨µµ·Î À¯ÀüÀÚ ¹ßÇöÀ» °ËÃâÇÒ ¼ö ÀÖ´Ù.
Intercalating dye-based qPCRÀº PCR ÁõÆø°úÁ¤ Áß¿¡ »ý¼ºµÇ´Â ÀÌÁß°¡´ÚÀÇ DNA¿¡ intercalating dye°¡ °áÇÕÇÏ´Â ¿ø¸®¸¦ ÀÌ¿ëÇÑ´Ù.
ÀϹÝÀûÀ¸·Î Dye-based qPCRÀº ¸¹Àº ¼öÀÇ »ùÇÿ¡¼­ ºü¸£°í °£´ÜÇÏ°Ô À¯ÀüÀÚ ¹ßÇöÀ» ºÐ¼®ÇÏ´Â °æ¿ì ÀûÇÕÇϸç, ¶óº§µÇÁö ¾ÊÀº ÀϹÝÀûÀÎ oligonucleotide¸¦ PCR ÁõÆø¿¡ »ç¿ëÇϹǷÎ, »ó´ëÀûÀ¸·Î Àú·ÅÇÏ°Ô ½ÇÇèÇÒ ¼ö ÀÖ´Ù.
RNA¸¦ ÀÌ¿ëÇÏ¿© ÇϳªÀÇ Æ©ºê¿¡¼­ ¿ªÀü»ç¹ÝÀÀÀ» ¸¶Ä£ ÈÄ, Æ©ºê¸¦ ¿Å°Ü qPCR ¹ÝÀÀÀ» ÁøÇàÇÏ´Â Two-step Real Time RT-PCR (¶Ç´Â two-step RT-qPCR)°ú ÇϳªÀÇ Æ©ºê¿¡¼­ ¿ªÀü»ç¹ÝÀÀ°ú qPCR ¹ÝÀÀÀ» ¿¬¼ÓÀûÀ¸·Î ÁøÇàÇÏ´Â One-step Real Time RT-PCR (¶Ç´Â one-step RT-qPCR)¿¡ ¸Â´Â ´Ù¾çÇÑ dye-based Real Time PCR ½Ã¾àÀ» ¼±ÅÃÇÒ ¼ö ÀÖ´Ù.

Dye-based qPCRÀÇ ÀåÁ¡
  • Rapid and simple protocol
  • More cost-effective than probes
  • Sensitive: 1,000X detection

Principles of green intercalating dye-based qPCR
Intercalating dye¸¦ ÀÌ¿ëÇÑ Çü±¤°ËÃâ
º» Real Time PCR ¹ýÀº ÀÌÁß°¡´ÚÀÇ DNA¿¡ °áÇÕÇßÀ» ¶§ Çü±¤À» ³ªÅ¸³»´Â DNA intercalator (¿¹, TB Green¢ç)À» »ç¿ëÇÑ´Ù. Çü±¤·®À» ¸ð´ÏÅ͸µÇÏ¿© ÁõÆø»ê¹°À» Á¤·®ÇÒ ¼ö ÀÖ´Ù (±×¸² 1). ÁõÆøÀÌ ¿Ï·áµÈ ÈÄ, melting curve ºÐ¼®À» ÅëÇØ PCR ÁõÆø»ê¹°ÀÌ Æ¯ÀÌÀûÀ¸·Î ÁõÆøµÇ¾ú´ÂÁö È®ÀÎÇÒ ¼ö ÀÖ´Ù. Takara Á¦Ç°Àº ¹ÝÀÀƯÀ̼º°ú °¨µµ¸¦ ±Ø´ëÈ­Çϱâ À§ÇØ hot start versionÀÇ TaKaRa Ex Taq¢â DNA polymerase¸¦ »ç¿ëÇϸç, À̸¦ ÅëÇØ ¹ÝÀÀ mixture Áغñ½Ã°£ ¶Ç´Â º» »çÀÌŬ °úÁ¤ Àü¿¡ mispriming ¶Ç´Â primer-dimerÀÇ Çü¼ºÀ¸·Î ¹ß»ýÇÒ ¼ö ÀÖ´Â ºñƯÀÌÀûÀÎ ÁõÆøÀ» ÃÖ¼ÒÈ­ÇÏ¿´´Ù.


±×¸² 1. Dye-based qPCR ¿ø¸®

Dye-based Real Time PCR Á¦Ç°
Takara qPCR premix¿¡´Â ³»¿­¼º Tli RNase H°¡ Æ÷ÇԵǾî ÀÖ¾î RNA¿¡¼­ cDNA·Î ¿ªÀü»çÇÑ ÈÄ, cDNA¿¡ °áÇյǾî ÀÖ´Â RNA¸¦ ¼±ÅÃÀûÀ¸·Î Á¦°ÅÇÏ¿© cDNA¿¡ PCR primer°¡ È¿À²ÀûÀ¸·Î °áÇÕÇÒ ¼ö ÀÖµµ·Ï µµ¿ÍÁØ´Ù.

±×¸² 2. ³»¿­¼º Tli RNase H °¡ Æ÷ÇԵǾî Á¤È®ÇÑ Á¤·® °¡´É

ÀϹÝÀûÀÎ »ùÇà Àû¿ë
    TB Green¢ç qPCRÀÇ Á¤¼®
        TB Green¢ç Premix Ex Taq¢â II (Tli RNaseH Plus) (Code RR820A)
    ½ÅÀå¼Óµµ 10ÃÊ °í¼Ó qPCR
        TB Green¢ç Fast qPCR Mix (Code RR430A)
    ÁõÆøÈ¿À² No.1
        TB Green¢ç Premix Ex Taq¢â (Tli RNaseH Plus) (Code RR420A)

Ư¼ö »ùÇà Àû¿ë
    ºñƯÀÌÀûÀÎ ÁõÆø, Primer dimer°¡ ³ªÅ¸³¯ ¶§
        TB Green¢ç Premix DimerEraser¢â (Perfect Real Time) (Code RR091A)
    gDNA, GC rich ¼­¿­ ÁõÆø¿¡ ÃÖÀû
        TB Green¢ç Premix Ex Taq¢â GC (Perfect Real Time) (Code RR071A)
    Crude »ùÇÃÀ̳ª GC ÇÔ·®ÀÌ 70% ³Ñ´Â »ùÇÿ¡ ÃÖÀû
        MightyAmp¢â for Real Time (TB Green¢ç Plus) (Code R075A)

TB Green¢ç Real Time PCR Premix Á¦Ç°¶óÀÎ

Code

Á¦Ç°¸í

¿ë·®*

RR820A

TB Green¢ç Premix Ex Taq¢â II (Tli RNaseH Plus)

200 ȸ

RR820L

TB Green¢ç Premix Ex Taq¢â II (Tli RNaseH Plus), Bulk

200 ȸ

RR82LR

TB Green¢ç Premix Ex Taq¢â II (Tli RNaseH Plus), ROX plus

200 ȸ

RR430A

TB Green¢ç Fast qPCR Mix

200 ȸ

RR420A

TB Green¢ç Premix Ex Taq¢â (Tli RNaseH Plus)

200 ȸ

RR420L

TB Green¢ç Premix Ex Taq¢â (Tli RNaseH Plus), Bulk

200 ȸ

RR42LR

TB Green¢ç Premix Ex Taq¢â (Tli RNaseH Plus), ROX plus

200 ȸ

RR071A

TB Green¢ç Premix Ex Taq¢â GC (Perfect Real Time)

200 ȸ

RR091A

TB Green¢ç Premix DimerEraser¢â (Perfect Real Time)

200 ȸ

R075A

MightyAmp¢â for Real Time (TB Green¢ç Plus)

200 ȸ

*50 §¡ ¹ÝÀÀ ½Ã


Citations
RR820A: Chen, Q., Gu, Y. & Liu, B. Expression and mechanism of action of the SARI tumor suppressor in prostate cancer. Int. J. Clin. Exp. Pathol. 8, 7953-60 (2015).
RR820A: Sato, Y. et al. Anks4b, a novel target of HNF4¥á protein, interacts with GRP78 protein and regulates endoplasmic reticulum stress-induced apoptosis in pancreatic ¥â-cells. J. Biol. Chem. 287, 23236-45 (2012).
RR420A: Sung, H. Y. et al. Amyloid beta-mediated hypomethylation of heme oxygenase 1 correlates with cognitive impairment in Alzheimer¡¯s disease. PLoS One 11, e0153156 (2016).