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Home > ÀüÁ¦Ç°º¸±â > cDNA ÇÕ¼º > Reverse Transcription (Takara) > PrimeScript¢â Reverse Transcriptase
Ź¿ùÇÑ ½ÅÀå´É·ÂÀ» °¡Áø ½Å°³³ä ¿ªÀü»ç È¿¼ÒÀÇ standard

PrimeScript¢â Reverse Transcriptase

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
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Takara
2680A
PrimeScript¢â Reverse Transcriptase
°ü·ÃÇмú ±¸¸ÅÇϱ⠻ùÇýÅû 
10,000 U
167,200¿ø 
209,000¿ø
°¡°ÝÇÒÀÎ
03.02 ~ 04.30
2680A_DS.v1901Da.pdf
Takara
2680B
PrimeScript¢â Reverse Transcriptase
°ü·ÃÇмú ±¸¸ÅÇϱâ
40,000 U
752,400¿ø 



³óµµ 200 U/§¡
Çü»ó
20 mM   Tris-HCl (pH7.8)
100 mM   NaCl
1 mM   EDTA
1 mM   DTT
50%   Glycerol(v/v)
º¸Á¸ -20¡É
5¡¿ PrimeScript Buffer (cDNA ÇÕ¼º¿ë)
250 mM   Tris-HCl (pH8.3)
375 mM   KCl
15 mM   MgCl2
Á¦Ç°¼³¸í
º» Á¦Ç°Àº M-MLV(Moloney Murine Leukemia Virus) À¯·¡ÀÇ Reverse Transcriptase¸¦ ±â¹ÝÀ¸·Î ÇÏ¿© ´ÙÄ«¶ó°¡ µ¶ÀÚÀûÀ¸·Î °³¹ßÇÑ ½Å±Ô ¿ªÀü»ç È¿¼ÒÀÌ´Ù. º» Á¦Ç°Àº ½ÅÀ强ÀÌ ¸Å¿ì ¶Ù¾î³ª ±ä cDNA¸¦ ÇÕ¼ºÇÒ ¼ö ÀÖ´Ù. ¶Ç, GC rich µî°ú °°ÀÌ cDNA ÇÕ¼ºÀÌ ¾î·Á¿î RNA¿¡
´ëÇØ¼­µµ RNA°¡ ºÐÇØµÉ ¿ì·Á°¡ ÀÖ´Â °í¿Â¿¡¼­ÀÇ ¹ÝÀÀÀº ÇÊ¿ä·Î ÇÏÁö ¾ÊÀ¸¸ç, Ç¥ÁØ ¿ªÀü»ç ¹ÝÀÀ ¿Âµµ(42¡É)¿¡¼­ È¿À²ÀûÀ¸·Î cDNA¸¦
ÇÕ¼ºÇÒ ¼ö ÀÖ´Ù. º» È¿¼Ò´Â ±ä cDNAÀÇ Á¶Á¦³ª full length cDNAÀÇ ºñÀ²ÀÌ ³ôÀº library Á¦ÀÛ¿¡ ÃÖÀûÀÌ´Ù.
PrimeScrip RTase ÀÌ¿ë 42¡É ¹ÝÀÀ¿Âµµ·Î, ¹é±×¶ó¿îµå°¡ ³·Àº °íǰÁúÀÇ cDNA ÇÕ¼º °¡´É

RNA Ladder (1~12kb)¸¦ ÁÖÇüÀ¸·Î, PrimeScript ¹× Ÿ»ç RTase¸¦ ÀÌ¿ëÇÏ¿© ¿ªÀü»ç ¹ÝÀÀ.
Ȱ¼ºÀÇ Á¤ÀÇ
Poly(rA)- oligo(dT)12-18 ¸¦ ÁÖÇü/ÇÁ¶óÀ̸ӷΠÇÏ¿© 37¡É, 10ºÐ°£ 1 nmolÀÇ [3H]dTTP¸¦ ÇÕ¼º±âÁú·Î »ç¿ëÇÏ´Â °ÍÀ» È¿¼Ò Ȱ¼º 1UÀ¸·Î ÇÑ´Ù..
Ȱ¼ºÃøÁ¤¿ë ¹ÝÀÀ¾× Á¶¼º
50 mM   Tris-HCl (pH8.3)
75 mM   KCl
8 mM   MgCl2
10 mM   DTT
20 ug/ml   (rA)n - (dT)12-18
0.5 mM   [3H]dTTP
0.1%   NP-40
1st-strand cDNA ÇÕ¼º½ÃÇè
Ç¥ÁØ ½ÇÇè¹ý(ÇÁ·ÎÅäÄÝ)¿¡ µû¶ó ·§Æ® ³ú À¯·¡ÀÇ Total RNA 100 ngÀ» ÁÖÇüÀ¸·Î ÇÏ¿© Oligo dT ÇÁ¶óÀÌ¸Ó 50 pmol ¹× º» È¿¼Ò 200 unitsÀ»
ÀÌ¿ëÇÏ¿© 42¡É¿¡¼­ 60ºÐ°£ 1st-strand cDNA ÇÕ¼º ¹ÝÀÀÀ» ÇÑ´Ù. ÀÌ cDNA ¹ÝÀÀ¾×À» ÁÖÇüÀ¸·Î PCR ÇÏ¿© ¾Æ°¡·ÎÁî °Ö¿¡ Àü±â ¿µµ¿ÇÑ
°á°ú 12 kbÀÇ size À§Ä¡¿¡¼­ ¸íÈ®ÇÑ ¹êµå¸¦ È®ÀÎÇÏ¿´´Ù.
¼øµµ
1. 200 UÀÇ º» È¿¼Ò¿Í 1§¶ÀÇ ¥ëDNA-Hind III ºÐÇØ¹°À» 37¡É, 1½Ã°£ ¹ÝÀÀ½ÃÄѵµ DNA Àü±â ¿µµ¿ ÆÐÅÏ¿¡ º¯È­°¡ ÀϾÁö ¾Ê´Â´Ù.
2. 200 UÀÇ º» È¿¼Ò¿Í 1§¶ÀÇ supercoiled pBR322 DNA¸¦ 37¡É, 1½Ã°£ ¹ÝÀÀ½ÃÄѵµ DNA Àü±â ¿µµ¿ ÆÐÅÏ¿¡ º¯È­°¡ ÀϾÁö ¾Ê´Â´Ù.
3. 200 UÀÇ º» È¿¼Ò¿Í 1§¶ÀÇ 23S ¹× 16S rRNA È¥ÇÕ¹°À» 37¡É, 1½Ã°£ ¹ÝÀÀ½ÃÄѵµ RNA Àü±â ¿µµ¿ ÆÐÅÏ¿¡ º¯È­°¡ ÀϾÁö ¾Ê´Â´Ù.
¿ëµµ
- 1st-strand cDNA ÇÕ¼º
- cDNA probe Á¶Á¦
- RT- PCR
Á¤È®µµ°¡ ¸Å¿ì ³ôÀº ¿ªÀü»çÈ¿¼Ò

Keyword : 2640A

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