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Home > ÀüÁ¦Ç°º¸±â > Stem Cell > iPS cell Á¦ÀÛ ¹× °ËÃâ > Retrovirus Titer Set (for Real Time PCR)
Virus »óûÀ¸·Î ºÎÅÍ Á÷Á¢ 4 ½Ã°£¿¡ RNA Titer¸¦ ÃøÁ¤

Retrovirus Titer Set (for Real Time PCR)

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Takara
6166
Retrovirus Titer Set (for Real Time PCR)
°ü·ÃÇмú ±¸¸ÅÇϱâ
100 ¹ÝÀÀºÐ
436,000¿ø  6166_e.v1904Da.pdf

³»¿ë (100 ¹ÝÀÀ ºÐ ; 25§¡ ¹ÝÀÀ°è*1)
Forward Titer Primer FRT-1 (10 pmol/§¡) 50 §¡
Reverse Titer primer RRT-1 (10 pmol/§¡) 50 §¡
RNA Control Template (5 ¡¿ 108 copies/§¡) 10 §¡
EASY Dilution (Real Time PCR ¿ë) 1 §¢ ¡¿ 2
DNase I (RNase Free) (5 U/§¡) 40 §¡
10 ¡¿ DNase I Buffer 50 §¡
RNase Inhibitor (porcine liver) (40 U/§¡) 10 §¡
*1 Real Time RT- PCR 100ȸ ±×¸®°í DNase I ó¸® 20ȸÀÓ.
º¸Á¸ -20¡É
RNA Control Template´Â ¹Ýº¹ÀûÀÎ µ¿°á À¶ÇØ¿¡ ÀÇÇÑ ºÐÇظ¦ ¸·±â À§ÇÏ¿© ¼Ò·® ºÐÁÖ ÈÄ Deep freezer (-70¡­-80¡É)¿¡¼­ ÀúÀåÇÏ´Â °ÍÀ» ÃßõÇÕ´Ï´Ù.
Á¦Ç°¼³¸í
º» Á¦Ç°Àº Real Time RT-PCRÀ» ÀÌ¿ëÇÏ¿© MLV (murine leukemia virus) À¯·¡ retrovirus vectorÀÇ RNA genome ·®À» ÃøÁ¤ÇÏ¿© ¿ª°¡(RNA titer)À» »êÃâÇÏ´Â Á¦Ç°ÀÌ´Ù.
º» Á¦Ç°À» One Step TB Green¢â PrimeScript RT-PCR Kit (Perfect Real Time) (Code RR066A/RR086A)°ú ÇÔ²² »ç¿ëÇÏ¿© ¹ÙÀÌ·¯½º »óµî¾×¿¡¼­ Á÷Á¢ RNA titer¸¦ ÃøÁ¤ÇÒ ¼ö ÀÖ°í, ¾à 4½Ã°£ ¾È¿¡ °á°ú¸¦ ¾òÀ» ¼ö ÀÖ´Ù. ¶ÇÇÑ, MLVÀÇ packaging signalºÎºÐÀ» ÁõÆø ´ë»óÀ¸·Î ÇÏ°í ÀÖ¾î °ÅÀÇ ¸ðµç MLV À¯·¡ RetrovirusÀÇ ¿ª°¡(RNA titer)¸¦ ÃøÁ¤ÇÒ ¼ö ÀÖ´Ù.
RetrovirusÀÇ °¨¿° ¿ª°¡¸¦ ÃøÁ¤ÇÒ °æ¿ì ÀϹÝÀûÀ¸·Î Ç¥Àû ¼¼Æ÷¿¡ Retrovirus¸¦ °¨¿°½ÃÅ°°í Çü±¤À̳ª ¾àÁ¦³»¼º µîÀ» ÀÌ¿ëÇÏ¿© °¨¿°È¿À²À» ÃøÁ¤ÇØ ¿ª°¡ (»ý¹°ÇÐÀû titer)À» »êÃâÇÏ´Â ¹æ¹ýÀÌ ÀÌ¿ëµÈ´Ù. ÀÌ ¹æ¹ýÀº ¼öÀÏÀÌ ¼Ò¿äµÇ¸ç, ¸¶Ä¿À¯ÀüÀÚ³ª ¾àÁ¦³»¼ºÀ¯ÀüÀÚ°¡ Æ÷ÇÔµÇÁö ¾ÊÀº Retrovirus vector¿¡´Â Àû¿ëÇÒ ¼ö ¾ø´Ù. ¶ÇÇÑ »ý¹°ÇÐÀû titer¸¦ ±âÁØÀ¸·Î MOI (multiplicity of infection)¸¦ ÃøÁ¤ÇØ ¼¼Æ÷¿¡ °¨¿°½Ãų °æ¿ì, ¿ª°¡ ÃøÁ¤¿¡ ¸çÄ¥ÀÌ ¼Ò¿äµÇ¹Ç·Î ÀÏ´Ü ¹ÙÀÌ·¯½º »óµî¾×À» µ¿°á º¸Á¸ÇÒ ÇÊ¿ä°¡ ÀÖ°í, µ¿°á À¶ÇØ¿¡ ÀÇÇØ ¹ÙÀÌ·¯½º ¿ª°¡°¡ ÀúÇ쵃 °¡´É¼ºÀÌ ÀÖÀ¸¹Ç·Î Ãʱâ ÃøÁ¤°ª°ú´Â ´Ù¸¥ MOIÀÇ ¹ÙÀÌ·¯½º°¡ ½ÇÇè¿¡ ÀÌ¿ëµÉ ¼ö ÀÖ´Ù.
º» Á¦Ç°Àº RetrovirusÀÇ RNA genome·®À» ÃøÁ¤Çϱ⠶§¹®¿¡ ¸¶Ä¿À¯ÀüÀÚ³ª ¾àÁ¦³»¼ºÀ¯ÀüÀÚ¿Í´Â °ü°è¾øÀÌ ¿ª°¡¸¦ ÃøÁ¤ÇÒ ¼ö ÀÖ´Ù. ¶ÇÇÑ ¾à 4½Ã°£ ¸¸¿¡ ÃøÁ¤ °á°ú¸¦ ¾òÀ» ¼ö ÀÖÀ¸¹Ç·Î ¹ÙÀÌ·¯½º »óµî¾×À» µ¿°áÇÏÁö ¾Ê°í ¿øÇÏ´Â MOI·Î °¨¿°½Ãų ¼ö ÀÖ´Ù. ÇÔ²² »ç¿ëÇÏ´Â One Step TB Green¢â PrimeScript RTPCR Kit (Perfect Real Time) Àº TB Green¢â °ËÃâ¿¡ ÀÇÇÑ 1 step Real Time RT-PCR Å°Æ®ÀÌ´Ù. ½ÅÀ强ÀÌ ¶Ù¾î³ª ´Ü½Ã°£¿¡ È¿À²ÀûÀ¸·Î cDNA ÇÕ¼ºÀÌ °¡´ÉÇÑ ¿ªÀü»ç È¿¼Ò PrimeScript ¿Í ´ÙÄ«¶óÀÇ ´ëÇ¥ÀûÀÎ PCR È¿¼ÒÀÎ TaKaRa Ex Taq HS¸¦ ÀÌ¿ëÇÏ¿© 1 step RT-PCR¿¡ ÃÖÀûÈ­ÇÏ°í ÀÖÀ¸¸ç ¾ÈÁ¤ÀûÀ¸·Î 1 step real time RT-PCRÀ» ½Ç½ÃÇÒ ¼ö ÀÖ´Ù.
ÀÌ¿Ü¿¡ ÇÊ¿äÇÑ ½Ã¾à
One Step TB Green¢â PrimeScript RT-PCR Kit (Perfect Real Time) (TaKaRa Code RR066A)
¶Ç´Â One Step TB Green¢â PrimeScript RT-PCR Kit II (Perfect Real Time) (TaKaRa Code RR086A)

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