- 1st strand cDNA ÇÕ¼º¿¡ ÇÊ¿äÇÑ ¸ðµç ½Ã¾àÀÌ Æ÷ÇÔµÈ premix
- 10~20ºÐÀÇ ¿ªÀü»ç¹ÝÀÀÀ¸·Î long fragment (13 kb) RNA ¿ªÀü»ç °¡´É
- RNA denaturation ºÒÇÊ¿ä - RNA ÷°¡ ÈÄ ¹Ù·Î RT ¹ÝÀÀ ½ÃÀÛ
- RTaseÀÇ ³»¿¼º Áõ°¡·Î ÇÊ¿ä ½Ã °í¿Â (55 ¡É)¿¡¼ ¹ÝÀÀ ÁøÇà
Á¦Ç°¼³¸í
PrimeScript™ IV 1st strand cDNA Synthesis Mix´Â total RNA ¶Ç´Â poly A+ RNA¿¡¼ 1st strand cDNA¸¦ ÇÕ¼ºÇϱâ À§ÇÑ Á¦Ç°ÀÌ´Ù. 1st strand cDNA ÇÕ¼º¿¡ ÇÊ¿äÇÑ ¿ªÀü»çÈ¿¼Ò PrimeScript™ IV RTase, RNase Inhibitor, Oligo dT primer, dNTP mixture ¹× ¹ÝÀÀ buffer°¡ premixµÇ¾î ÀÖ¾î template RNA¿Í D.W¸¦ ÷°¡ÇÏ´Â °Í ¸¸À¸·Î ½±°í ºü¸£°Ô ¿ªÀü»ç ¹ÝÀÀÀ» ÁøÇàÇÒ ¼ö ÀÖ´Ù.
º» Á¦Ç°Àº ¾Ç¼¼¼¸® ´Ü¹éÁúÀÌ Æ÷ÇÔµÇ¾î ±âÁ¸ ¿ªÀü»ç ¹ÝÀÀ¿¡ ÇÊ¿ä·Î Çß´ø RNA denaturation °úÁ¤À» »ý·«ÇÏ°í, RNAÀÇ ¼Õ»ó ¾øÀÌ ´õ °£´ÜÇÏ°Ô cDNA ÇÕ¼ºÀÌ °¡´ÉÇØÁ³´Ù.
½Å±Ô ¿ªÀü»ç È¿¼ÒÀÎ PrimeScript™ IV RTase´Â ±âÁ¸ Á¦Ç°º¸´Ù ªÀº ½Ã°£¿¡ cDNA¸¦ ÇÕ¼ºÇÒ ¼ö ÀÖ°í, ³»¿¼ºÀÌ Çâ»óµÇ¾î gene specific primer¸¦ »ç¿ëÇÏ´Â °æ¿ì ³ôÀº ¿Âµµ¿¡¼ ³ôÀº ƯÀ̼ºÀ¸·Î cDNA¸¦ ÇÕ¼ºÇÒ ¼ö ÀÖ´Ù.
º» Á¦Ç°À¸·Î ÇÕ¼ºµÈ 1st strand cDNA´Â 2nd strand ÇÕ¼º, hybridization, PCR ÁõÆø, full length cDNA library Á¦ÀÛ µî °íÇ°ÁúÀÇ full length cDNA°¡ ÇÊ¿äÇÑ ÈļӽÇÇè¿¡ ÀûÇÕÇÏ´Ù. ¶ÇÇÑ Á¦Ç° ³»¿¡ ±¸¼ºµÇ¾î ÀÖ´Â Random 6 mers¸¦ »ç¿ëÇÏ¿© poly A+°¡ ¾ø´Â RNA·ÎºÎÅÍ cDNA ÇÕ¼ºÀÌ °¡´ÉÇÏ°í, À¯ÀüÀÚ ¹ßÇö ºÐ¼®À» À§ÇÑ Real Time PCR¿ë cDNA¸¦ ÇÕ¼ºÇÒ ¼ö ÀÖ´Ù.
³»¿ë (Code 6215A, 20 §¡ ¹ÝÀÀ ½Ã 50ȸ ºÐ·®)
5× PrimeScript™ IV cDNA Synthesis Mix *1
|
200 §¡
|
Random 6 mers (50 μM) *2
|
100 §¡
|
RNase Free dH2O
|
1 §¢ x2
|
*1 PrimeScript™ IV RTase, RNase Inhibitor, Oligo dT Primer, dNTP Mixture ¹× ¹ÝÀÀ buffer (Mg2+ ÇÔÀ¯)À» Æ÷ÇÔÇÑ´Ù.
*2 poly A
+À» Æ÷ÇÔÇÏÁö ¾Ê´Â RNAÀÇ cDNA ÇÕ¼º ¶Ç´Â Real Time PCR¿ë cDNA ÇÕ¼º µîÀÇ °æ¿ì ¹ÝÀÀ¾×¿¡ Ãß°¡ÇÑ´Ù.
º¸Á¸ -20¡É
½ÇÇè ¿¹½Ã
- »ç¿ëÀÚ ¸Þ´º¾ó Âü°í
°ü·ÃÁ¦Ç°
- High Fidelity, Full Length cDNA ÇÕ¼º °ü·ÃÁ¦Ç°
- gDNA Eraser spin columnÀ¸·Î ´Ù¾çÇÑ »ùÇÿ¡¼ gDNA ¿À¿° ¾ø´Â RNA ÃßÃâ TaKaRa MiniBEST Universal RNA Extraction Kit (Code 9767A)
- ±Ø°ÀÇ ÆíÀǼº »ó¿Âº¸°ü µ¿°á°ÇÁ¶ premix RTase RNA to cDNA EcoDry Premix (Code 639541)
- Quick & Effective PCR ºü¸¥ ¹ÝÀÀ¼Óµµ, 2X premix, dye plus PrimeSTAR® GXL Premix Fast, Dye plus (Code R052A)
- ½ÇÇ躰 PCR È¿¼Ò ¼±ÅÃÇϱâ