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Home > ÀüÁ¦Ç°º¸±â > cDNA ÇÕ¼º > Reverse Transcription (Clontech) > SMARTScribe Reverse Transcriptase

SMARTScribe Reverse Transcriptase

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Clontech
639536
SMARTScribe Reverse Transcriptase
°ü·ÃÇмú ±¸¸ÅÇϱ⠻ùÇýÅû 
40 rxns
147,200¿ø 
184,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
x33065
Clontech
639537
SMARTScribe Reverse Transcriptase
°ü·ÃÇмú ±¸¸ÅÇϱâ
100 rxns
318,400¿ø 
398,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Clontech
639538
SMARTScribe Reverse Transcriptase
°ü·ÃÇмú ±¸¸ÅÇϱâ
400 rxns
1,142,400¿ø 
1,428,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.27
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â

  • ÃÖ´ë 14.7 kbÀÇ full-length cDNA ÇÕ¼º
  • ÀûÀº ¾çÀÇ »ùÇà Àû¿ë °¡´É
  • ÁÖÇü RNAÀÇ cmoplexity À¯Áö
  • Àü»çüÀÇ 5¡¯ end ºÐ¼® °¡´É
Á¦Ç°¼³¸í
SMARTScribe¢â Reverse Transcriptase Àº ÆíÇ⼺(bias)ÀÌ ¾øÀÌ cDNA¸¦ ÇÕ¼ºÇÏ´Â °í¼º´É È¿¼Ò·Î, ¸ðµç RNA Àü»çü¿¡¼­ ÁõÆø ¹× ¶óÀ̺귯¸® ±¸ÃàÀÌ °¡´ÉÇÏ´Ù. SMARTScribe¢â RTase´Â ÁÖÇü RNA »ùÇÃÀÇ ºñÀ²À» º¸Á¸Çϸ鼭, ±ä Àü»ç¹°¿¡¼­ full-length cDNA (14.7 kb ±îÁö)¸¦ ÇÕ¼ºÇÏ´Â °³¼±µÈ Moloney Murine Leukemia Virus reverse transcriptase (MMLV RTase)ÀÌ´Ù. ´ç»ç¸¸ÀÇ Á¤Á¦ °øÁ¤°ú ¾ö°ÝÇÑ Ç°Áú °ü¸® ±âÁØÀ» ÅëÇØ SMARTScribe¢â RTase¿¡ ¿À¿°À» ÀÏÀ¸Å³ ¼ö ÀÖ´Â nucleases¸¦ ¸ðµÎ Á¦°ÅÇÏ¿´À½À» È®ÀÎÇÏ¿´´Ù.

º» È¿¼Ò´Â ´Ù¾çÇÑ ÀÀ¿ë ºÐ¾ß¿¡ Àû¿ë °¡´ÉÇØ ´ç»çÀÇ SMART¢ç technology¸¦ ±â¹ÝÀ¸·Î ÇÏ´Â NGS Á¦Ç°°ú ÇÔ²² »ç¿ëµÇµµ·Ï Ưº° Á¦À۵Ǿú´Ù.

±×¸² 1. SMARTScribe¢â RTaseÀÇ ¶Ù¾î³­ °¨µµ.
SMARTScribe¢â RTase ¹× ÇÕ¼º RNA¸¦ »ç¿ëÇÏ¿© RT-PCRÀ» ÁøÇàÇÏ¿´´Ù. ÇÕ¼º RNA¸¦ 10¹è¾¿ serial dilutionÇÏ¿© »ùÇà ´ç 105 - 10 °³ copies¸¦ ¾ò¾ú´Ù. ±×·± ´ÙÀ½ Èñ¼®ÇÑ ÁÖÇüÀ» HeLa ¼¼Æ÷ÀÇ Total RNA 50 ng¿¡ ÷°¡ÇÏ¿© º¹À⼺À» Áõ°¡½ÃÄ×´Ù. ÀÌ·¸°Ô ÁغñÇÑ »ùÇðú SMARTScribe¢â RTase·Î ÇÕ¼ºÇÑ first-strand cDNA¸¦ Advantage¢ç 2 DNA Polymerase Mix¿Í ƯÀÌÀûÀÎ primer¸¦ »ç¿ëÇÏ¿© 350 bp Å©±âÀÇ ÁõÆø»ê¹°ÀÌ µÇµµ·Ï 36 cycleÀÇ PCR ¹ÝÀÀÀ» ÁøÇàÇØ ÁõÆøÇß´Ù. PCR ¹ÝÀÀ»ê¹°À» ¾Æ°¡·Î½º °Ö Àü±â ¿µµ¿À¸·Î ºÐ¼®ÇÑ °á°ú, SMARTScribe¢â RTase´Â ºÒ°ú 10 copiesÀÇ ÇÕ¼º RNA¿¡¼­µµ ´ÜÀÏ °¡´Ú cDNA¸¦ »ý¼ºÇÒ ¼ö ÀÖ¾ú´Ù. ÀÌ´Â SMARTScribe¢â RTase°¡ ÁÖÇü RNA template¿¡ ÁÁÀº °¨µµ¸¦ º¸Àδٴ °ÍÀ» ÀǹÌÇÑ´Ù. (Lane M: 100 bp DNA size marker)


±×¸² 2. Ÿ»ç Á¦Ç°º¸´Ù ´õ sensitiveÇÑ SMARTScribe¢â RTase.
Human Universal Total RNA¸¦ 10¹è¾¿ serial dilution(1 §¶¿¡¼­ 0.1 pg±îÁö)ÇÏ¿© SMARTScribe¢â RTase(ÆгΠA), °æÀï»ç P(ÆгΠB) ¶Ç´Â °æÀï»ç Q(ÆгΠC)¸¦ »ç¿ëÇÏ¿© °¢ È¿¼Ò Á¦Á¶¾÷üÀÇ ¸Å´º¾ó¿¡ µû¶ó ¿ªÀü»ç¹ÝÀÀÀ» ½Ç½ÃÇÏ¿´´Ù. ÀÌÈÄ °¢ RT ¹ÝÀÀÀÇ 1 §¡·Î ¥â-actin À¯ÀüÀÚÀÇ 5' ¸»´Ü¿¡ ƯÀÌÀûÀÎ primer¸¦ »ç¿ëÇØ PCR ÇÏ¿´°í, ¾Æ°¡·Î½º °Ö Àü±â¿µµ¿À¸·Î ºÐ¼®ÇÑ °á°ú, SMARTScribe¢â RTase´Â ¸ðµç template ³óµµ¿¡¼­ ¶Ù¾î³­ °¨µµ¸¦ ³ªÅ¸³Â´Ù.
(°¢ ¹ÝÀÀ¿¡ »ç¿ëµÈ template RNAÀÇ ¾ç. Lane 1: 1 §¶, Lane 2: 100 ng, Lane 3: 10 ng, Lane 4: 1 ng, Lane 5: 100 pg, Lane 6: 10 pg, Lane 7: 1 pg, Lane 8: 0.1 pg.)
³»¿ë
  • 100 U/§¡SMARTScribe¢â Reverse Transcriptase
  • 5X First-Strand Buffer
  • 20 mM DTT
º¸Á¸
- 20 ¡É
°ü·Ã Á¦Ç°
Applications
- RT-PCR
- Real-time PCR/qPCR
- Gene expression analysis
- Cloning
- Library construction
- Microarray

Keyword :

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