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Home > ÀüÁ¦Ç°º¸±â > Real Time PCR > Cell direct qPCR > Whole Transcriptome ÁõÆø½Ã¾à
1 °³ÀÇ ¼¼Æ÷·ÎºÎÅÍ template cDNA¸¦ ÁõÆø °¡´É

Whole Transcriptome ÁõÆø½Ã¾à

-

Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Takara
3734
CellAmp¢â Whole Transcriptome Amplification Kit (Real Time) Ver.2
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
100 ȸ
(5 §¡ ¹ÝÀÀ°è)
625,600¿ø 
782,000¿ø
°¡°ÝÇÒÀÎ
09.02 ~ 10.31
3734_e.v2201Da.pdf

  • 1~1,000°³ Á¤µµ ¼¼Æ÷ ȤÀº 20 pg~20 ngÀÇ total RNA¿¡¼­ È¿À²ÀûÀ¸·Î cDNA ÁõÆø °¡´É
  • ¹Ì·® »ùÇÿ¡¼­µµ ´Ù¾çÇÑ À¯ÀüÀÚÀÇ Real Time PCR Çؼ® °¡´É
  • ÇÁ·ÎÅäÄÝ ÃÖÀûÈ­ ÅëÇØ Àú¹ßÇö À¯ÀüÀÚ±îÁö È¿À²ÀûÀ¸·Î ÁõÆø °¡´É
Á¦Ç° ¼³¸í
º» Á¦Ç°Àº ¼Ò¼öÀÇ ¼¼Æ÷(1~1000°³)¸¦ ÀÌ¿ëÇÏ¿© ¹Ù·Î cDNA¸¦ ÁõÆøÇÏ´Â Á¦Ç°ÀÌ´Ù. ÁõÆøµÈ cDNA´Â Real Time PCR ÁÖÇüÀ¸·Î »ç¿ëÇÒ ¼ö ÀÖ´Ù. ÀϹÝÀûÀ¸·Î ¹Ì·®ÀÇ ÇÙ»êÀ» ÀÌ¿ëÇÒ ¶§, Á¤Á¦¿¡ ÀÇÇÑ ¼Ò½ÇÀÌ ¹®Á¦°¡ µÇÁö¸¸, ÀÌ Á¦Ç°À» ÀÌ¿ëÇϸé RNA³ª cDNA Á¤Á¦¾øÀÌ È¿À²ÀûÀ¸·Î cDNA¸¦ ÁõÆøÇÒ ¼ö ÀÖ´Ù. º» Á¦Ç°¿¡¼­´Â ¿ì¼± ¼¼Æ÷¸¦ ¿ëÇØÇÏ¿©, dT-adaptor primer(RT dT Primer 2)¸¦ ÀÌ¿ëÇÑ ¿ªÀü»ç ¹ÝÀÀ ÈÄ, mRNA·ÎºÎÅÍ cDNA¸¦ ÇÕ¼ºÇÑ´Ù. ±× ´ÙÀ½¿¡ ÇÕ¼ºÇÑ cDNA¿¡ TdT·Î dA-tail¸¦ ºÎ°¡ÇÑ ÈÄ, ÀÌ°ÍÀ» ÁÖÇüÀ¸·Î ÇÏ¿© PCR cycle ¼ö¸¦ ÁÙ¿© cDNA¸¦ ÁõÆøÇÑ´Ù.
º» Á¦Ç°Àº ¼¼Æ÷¿¡¼­ Á÷Á¢ cDNA ÁõÆøÇÒ ¼öµµ ÀÖ°í, ¹Ì·®ÀÇ RNA(20 pg~20 ng)¿¡¼­ cDNA ÁõÆøÇÒ ¼öµµ ÀÖ´Ù.
Ver.2¿¡¼­´Â Exonuclease Ió¸® °úÁ¤À» Ãß°¡ÇÏ¿©, ±âÁ¸ Á¦Ç°¿¡ ºñÇØ ÇÁ¶óÀÌ¸Ó À¯·¡ÀÇ ºñƯÀÌÀû ÁõÆøÀ» ¾ïÁ¦ÇÏ¿© ¹ßÇö·®ÀÇ ÀûÀº À¯ÀüÀÚÀÇ ÁõÆø È¿À²À» Çâ»ó½ÃÄ×´Ù.
³»¿ë
(5 ¥ìl¹ÝÀÀ½Ã, 100ȸºÐ)
1. Lysis Buffer£¨4¡¿£© 150 ¥ìl
2. Recombinant RNase Inhibitor£¨40 U/¥ìl£© 30 ¥ìl
3. RT dT Primer 2 12 ¥ìl
4. dNTP Mixture£¨2.5 mM each£© 12 ¥ìl
5. MgCl2£¨22.5 mM£© 36 ¥ìl
6. RT Enzyme Mix 2*1 36 ¥ìl
7. Exonuclease I£¨5 U/¥ìl£© 72 ¥ìl
8. TdT Buffer£¨5¡¿£© 144 ¥ìl
9. dATP£¨90 mM£© 24 ¥ìl
10. TdT Enzyme Mix*2 54 ¥ìl
11. PCR Primer Mix 2 300 ¥ìl
12. RNase Free dH2O 1 ml

*1 PrimeScript Reverse Transcriptase¡¢RNase Inhibitor Æ÷ÇÔ.
*2 Terminal Deoxynucleotidyl Transferase¡¢RNase H Æ÷ÇÔ.
º¸Á¸
- 20 ¡É
º» Á¦Ç° »ç¿ë½Ã ÇÊ¿äÇÑ ½Ã¾à ¹× ±â±â (ÁÖ¿ä Ç°¸ñ)
PCR ½Ã¾à
- TaKaRa Ex Taq Hot Start Version(Code RR006A/B)
1.5 ml microtube
0.2 ml microtube
Micropipette ¹× tip
PCR Thermal Cycler
- TaKaRa PCR Thermal Cycler Dice Gradient/Standard(Á¦Ç° ÄÚµå TP600/TP650) µî
½ÇÇè°úÁ¤

Real Time RT-PCR¿ë primer Á¦ÀÛ¼­ºñ½º¡¸Perfect Real Time Support System¡¹
Perfect Real Time Support SystemÀº intercalator ¹ý(SYBR Green I°ËÃâ)À» ÀÌ¿ëÇÑ Real Time RT-PCR¿ëÀ¸·Î ÃÖÀûÈ­µÈ ÇÁ¶óÀ̸Ӹ¦
°Ë»ö¡¤¿Â¶óÀÎ ÁÖ¹®Çϱâ À§ÇÑ ½Ã½ºÅÛÀÌ´Ù.
Human, mouse, rat, dog, chicken, bovineÀÇ RefSeq µî·Ï À¯ÀüÀÚ¿¡ ´ëÇÑ Àü¹ÝÀûÀÎ Real Time RT-PCR¿ë ÇÁ¶óÀÌ¸Ó ¼³°è°¡ °¡´ÉÇÏ´Ù.
Ÿ°Ù À¯ÀüÀÚ¸¦ °Ë»öÇÏ¿©, ¿øÇÏ´Â ÇÁ¶óÀÌ¸Ó ¼¼Æ®¸¦ ¼±ÅÃÇÏ¸é °£´ÜÇÏ°Ô ¿Â¶óÀÎÀ¸·Î ÁÖ¹® °¡´ÉÇÏ´Ù. ¹ø°Å·Î¿î ÇÁ¶óÀÌ¸Ó ¼³°è¿Í ¹ÝÀÀ Á¶°Ç ÃÖÀûÈ­°¡ ºÒÇÊ¿ä(TAKARA Real Time PCR reagent¿¡ ÃÖÀûÈ­) Çϸç, siRNAÀÇ È¿°úÀÇ È®ÀÎ, DNA Microarray Çؼ® °á°ú °ËÁõ µî À¯ÀüÀÚ ¹ßÇö Á¤·®À̳ª À¯ÀüÀÚ °ËÃâ¿¡ °£ÆíÇÏ°Ô »ç¿ëÇÒ ¼ö ÀÖ´Ù.

[Real Time Support System ¹Ù·Î°¡±â]

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1 °³ÀÇ ¼¼Æ÷·ÎºÎÅÍ template cDNA¸¦ ÁõÆø °¡´É
Whole Transcriptome ÁõÆø½Ã¾à