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Gene transfer (virus)
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Retrovirus
> qRT-PCR titer ÃøÁ¤
4½Ã°£ À̳»¿¡ Recombinant RetrovirusÀÇ RNA titer¸¦ ÃøÁ¤
qRT-PCR titer ÃøÁ¤
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Clontech
631453
Retro-X qRT-PCR Titration Kit
200 ȸ
(Package)
856,000¿ø
1,070,000¿ø
11.01 ~ 12.27
,
,
Product index
Adeno-Associated Virus (AA..
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4½Ã°£ À̳»¿¡ Recombinant RetrovirusÀÇ RNA titer¸¦ ÃøÁ¤
ÇϷ縸¿¡ ¹ÙÀÌ·¯½º ȸ¼öºÎÅÍ titer ÃøÁ¤°ú transduction °¡´É
MMLV ±â¹ÝÀÇ Retroviral vector¿¡ »ç¿ë °¡´É (* MSCV ±â¹ÝÀÇ Retrovirus¿¡´Â »ç¿ë ºÒ°¡)
Á¤È®ÇÑ titer¸¦ ¾òÀ» ¼ö ÀÖÀ¸¸ç ¾ÈÁ¤µÈ ½ÇÇèÀÌ °¡´É
Retro-X qRT-PCR Titration Kit¸¦ ÀÌ¿ëÇϸé recombinant retrovirusÀÇ titer°ªÀ» ½Å¼Ó, °£ÆíÇÏ°í Á¤È®ÇÏ°Ô ÃøÁ¤ ÇÒ ¼ö ÀÖ´Ù. °£´ÜÇÏ°Ô RNA Á¤Á¦¸¦ ½Ç½Ã ÇÑ ÈÄ, One-Step qRT-PCRÀ» ÀÌ¿ëÇÏ¿© ½Ã·á¿Í control RNA (kit¿¡ Á¦°ø)¸¦ serial dilutionÇÏ°í, Èñ¼®µÈ ½Ã·áÀÇ copy¼ö¸¦ standard curveÀÇ Ct °ª°ú ºñ±³ÇÏ¿© °£´ÜÇÏ°Ô ¿ª°¡¸¦ °áÁ¤ÇÒ ¼ö ÀÖ´Ù. (±×¸²1).
±×¸²1. Flowchart of the procedure used for titrating retroviral supernatants with the qRT-PCR titration kits.
´Ü½Ã°£ÀÇRNA titerÃøÁ¤¿¡ ÀÇÇØ ÀçÇö¼ºÀÌ ³ôÀº °¨¿° ½ÇÇèÀÌ °¡´É
°¨¿°¼ºÀ» ±âº»À¸·Î ÇÑ Ç¥ÁØÀûÀÎ titer ÃøÁ¤¹ý¿¡´Â Çü±¤¸¶Ä¿, ¾àÁ¦³»¼º, ¼¼Æ÷¿°»öÀ» ÀÌ¿ëÇÑ ¹æ¹ýÀÌ ÀÖÁö¸¸, titer¸¦ ÃøÁ¤Çϴµ¥ 2~10ÀÏÁ¤µµ ¼Ò¿äµÈ´Ù.(±×¸²2). °¡Àå ½Å¼ÓÇÏ°Ô ¿ª°¡¸¦ ÃøÁ¤ ÇÒ¼ö ÀÖ´Â ¹æ¹ýÀÎ flow cytometery¸¦ ÀÌ¿ëÇÏ´õ¶óµµ 48½Ã°£ ¹Ì¸¸¿¡¼´Â Á¤È®ÇÏ°Ô ÃøÁ¤ ÇÒ ¼ö ¾ø´Ù.
ÀÌ·¯ÇÑ Ç¥ÁØÀûÀÎ titer ÃøÁ¤¹ýÀº Àå½Ã°£À» ÇÊ¿ä·Î Çϱ⠶§¹®¿¡ ¹Ýº¹ÀûÀÎ µ¿°á Çص¿ ¶Ç´Â Àå±â°£ º¸°ü ÇÒ ¼ö¹Û¿¡ ¾ø´Ù. ±× »çÀÌ¿¡ ¹ÙÀÌ·¯½ºÀÇ °¨¿°¼ºÀÌ ÀúÇÏ µÉ ¼ö ÀÖ´Ù. ÇÏÁö¸¸ ÀúÇϵǴ Á¤µµ´Â ÃøÁ¤ ÇÒ ¼ö ¾ø±â ¶§¹®¿¡ ¸¹Àº °æ¿ì ¸îÀÏÀÌ Áö³ ÃßÁ¤ titer¸¦ ÀÌ¿ëÇÏ¿© °¨¿°¼º°ú MOI(multiplicity of infection)ÀÇ »êÃâ¿¡ »ç¿ëÇÏ°Ô µÈ´Ù.
±×¸²2. Lenti-X and Retro-X qRT-PCR Titration Kits offer the fastest titration methods. Standard transduction-based titration methods require up to 10 days to complete. The Lenti-X and Retro-X qRT-PCR Titration Kits require only 4 hr, which enables virus harvest, titration, and infection of target cells to all be performed on the same day.
Ç¥ÁØÀûÀÎ ÇüÁú µµÀÔ¿¡ ÀÇÇÑ titerÃøÁ¤¹ýÀº ÃÖÀå 10ÀÏÀÌ ÇÊ¿äÇÏ´Ù. ÇÏÁö¸¸ Retro-X qRT-PCR Titration Kit¸¦ ÀÌ¿ëÇÏ¸é ºÒ°¡ 4½Ã°£ÀÌ ¼Ò¿äµÇ±â ¶§¹®¿¡ ¹ÙÀÌ·¯½ºÀÇ È¸¼ö, ¿ª°¡ ÃøÁ¤ ¹× Ÿ°Ù ¼¼Æ÷¿¡ °¨¿°±îÁö ÇÏ·ç ¸¸¿¡ ½Ç½Ã ÇÒ ¼ö ÀÖ´Ù. ½Å¼ÓÇÏ°Ô ¹ÙÀÌ·¯½ºÀÇ ¿ª°¡¸¦ ÃøÁ¤ÇϹǷμ ¹ÙÀÌ·¯½º ¿ª°¡ÀÇ ÀúÇϸ¦ ¹æÁöÇϸç, ÀǵµÇÑ´ë·Î MOI ¹× ÇüÁúµµÀÔÀ» ÇÒ ¼ö ÀÖÀ¸¸ç, ÀçÇö¼º ³ôÀº °á°ú¸¦ ¾òÀ» ¼ö ÀÖ´Ù.
±×¸² 3. Expression increases with higher MOIs. Histogram plots of NIH 3T3 cells transduced with increasing amounts of supernatant containing an amphotropic retrovirus that expresses the ZsGreen1 fluorescent protein. Greater amounts of virus increase the number of infected cells expressing ZsGreen1. Panels A-C. MOIs of 0.3, 3, and 30, respectively, resulted in 8%, 50%, and 97% ZsGreen1-positive cells having a fluorescence intensity greater than 10 (M1 population).
qRT-PCR¿ª±â ÃøÁ¤¹ý
º» Á¦Ç°Àº 200 ȸÀÇ qRT-PCR¹ÝÀÀÀ» ¼öÇàÇÒ ¼ö ÀÖ´Â ¹ÝÀÀ ½Ã¾à°ú ¹ßÇö µÇ¾îÁö´Â ¼¿ (cDNA³ª shRNA)°ú »ó°ü¾øÀÌ HIV-1±â¹ÝÀÇ lentivirus ¶Ç´Â MMLV ±â¹ÝÀÇ retroivirus vector ¾çÂÊ ¸ðµÎ¿¡ »ç¿ëÇÒ ¼ö ÀÖ´Â ¿ª°¡ ÃøÁ¤¿ë primer set·Î ±¸¼ºµÇ¾î ÀÖ´Ù. Vector°¡ ´Ù¾çÇصµ ±âº» °ñ°Ý ±¸Á¶°¡ µ¿ÀÏÇÏ¿© °ËÃâ¹æ¹ýÀ̳ª selectable marker¸¦ º¯°æÇÏÁö ¾Ê°íµµ Çѹø¿¡ ¿ª°¡¸¦ ÃøÁ¤ÇÒ ¼ö ÀÖÀ¸¸ç, viral RNA Á¤Á¦¿Í DNaseI ó¸® ´Ü°è¸¦ ÅëÇÏ¿© ÀÜÁ¸ °¡´É¼ºÀÌ ÀÖ´Â plasmid DNA¸¦ Á¦°ÅÇÏ¿© ÀϽÃÀû transfectionÀ¸·ÎºÎÅÍ »ý¼ºµÈ ¹ÙÀÌ·¯½ºµéÀ» ÃÖ¼ÒÇÑÀÇ ½Ã·á¸¦ »ç¿ëÇÏ¿©µµ Á¤È®ÇÏ°Ô Á¤·®ÇÒ ¼ö°¡ ÀÖ´Ù.
±×¸²4. The qRT-PCR retroviral titration kit has a wide dynamic range. Panel A. Amplification plots of a qRT-PCR titration of the Retro-X RNA Control Template from 10
9
-10
4
copies using the Retro-X qRT-PCR Titration Kit. The assay shows a dynamic range of at least six orders of magnitude with low No-Template-Control (NTC) background. Panel B. A standard curve of the titration from Panel A demonstrates a strong linear correlation between Ct and the RNA copy number (log scale), with R
2
= 1.000 and a PCR efficiency of 107.4%.
Components & Storage Conditions
°¢ Á¦Ç° ±¸¼º¹°°ú º¸°üÁ¶°ÇÀº Certificate of Analysis ¸¦ ÂüÁ¶ÇϽʽÿÀ.
Keyword :
Retrovirus
Retro-X ¼±Åà °¡À̵å
RetroNectin¢çÀÇ ´Ù¾çÇÑ ÃֽŠapplication
Packaging Cell £¦ ½Ã½ºÅÛ
Retrovirus vector £¦ ½Ã½ºÅÛ
Çü±¤´Ü¹éÁú Retrovirus vector
MSCV Retrovirus ½Ã½ºÅÛ
Retro-X Q vector
À¯ÀüÀÚ µµÀÔ È¿À²°ú ¹ßÇö ±ÕÇü¼ºÀÌ ÁÁ¾Æ Ãʺ¸ ..
pDON-5 DNA
°íÈ¿À²·Î À¯ÀüÀÚ µµÀÔ °¡´ÉÇÑ Retrovirus Vecto..
pDON-AI-2 DNA
°íÈ¿À²·Î À¯ÀüÀÚ µµÀÔÀÌ °¡´ÉÇÑ Retrovirus vec..
pMEI-5 DNA
ÀϽÃÀûÀ¸·Î ³ôÀº titerÀÇ Recombinant Retrovir..
Retrovirus Packaging Kit
Tet À¯µµ¹ßÇö ½Ã½ºÅÛ (Tet-inducible-two vectors)
Single vectorÀÇ Retro-viral Tet À¯µµ¹ßÇö ½Ã..
Tet One Inducible System (one vector)
4½Ã°£ À̳»¿¡ Recombinant RetrovirusÀÇ RNA ti..
qRT-PCR titer ÃøÁ¤
Virus »óû¾×¿¡¼ Á÷Á¢ 4 ½Ã°£¿¡ RNA titer ¸¦ ..
Titer Set (for Real Time PCR)
Retrovirus ³óÃà
Receptor Boosters (Ecotropic and Amphotropic)
Cycling Probe·Î ½Å¼ÓÇÏ°Ô Provirus Copy ¼ö¸¦ ..
Provirus Copy ¼ö ÃøÁ¤
Integration Site Analysis Kit