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Home > ÀüÁ¦Ç°º¸±â > Gene transfer (virus) > Lentivirus > Provirus Quantitation Kit
¸ñÀû ¼¼Æ÷ÀÇ À¯ÀüÀÚ¿¡ »ðÀÔµÈ Provirus copy number Á¤·®

Provirus Quantitation Kit

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Clontech
631239
Lenti-X¢â Provirus Quantitation Kit
°ü·ÃÇмú ±¸¸ÅÇϱâ
200 ȸ
(Package)
1,029,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x101679, x32865, x100175

  • ½ÇÇè °£ÀÇ transgene expression levelÀ» ¿¹Ãø °¡´É
  • ¿©·¯ ¼¼Æ÷ ÁÖ°£ÀÇ ÇüÁúµµÀÔ µ¥ÀÌÅÍ ºñ±³
  • ½ÇÇè °£ ¶Ç´Â ´Ù¸¥ ¼¼Æ÷ÁÖ °£ÀÇ ÀÏ°üµÈ copy number È®º¸
  • Genomic DNA purification kit 10ȸºÐ, TB Green qPCR premix 200ȸºÐ Æ÷ÇÔ

¸ñÀû¼¼Æ÷ÀÇ nuclear DNA¿¡ »ðÀÔµÈ lentivirusÀÇ ¾çÀ» Á¤·®ÇÔÀ¸·Î½á, Lentiviral supernatant stock ³»¿¡ Á¸ÀçÇÏ´Â ½ÇÁ¦ ¿ª°¡ (real titer, effective titer)¿Í MOI (Multiplicity of infection)¸¦ º¸´Ù Á¤È®ÇÏ°Ô ÃøÁ¤ÇÒ ¼ö ÀÖ´Ù. À̸¦ ÅëÇÏ¿© ¾ó¸¶³ª ¸¹Àº ¹ÙÀÌ·¯½º À¯Àüü (viral genome)ÀÌ ¸ñÀû ¼¼Æ÷¿¡ »ðÀÔÀÌ µÇ¾ú´ÂÁö¸¦ ¿¹ÃøÇÒ ¼ö ÀÖ´Ù.
Lenti-X¢â Provirus Quantitation Kit´Â tranduced cell ³»ÀÇ »ðÀÔµÈ ·»Æ¼¹ÙÀÌ·¯½º (integrated lentivirus (provirus))ÀÇ copy number¸¦ ºü¸£°Ô Á¤·®ÇÒ ¼ö ÀÖ´Â Á¦Ç°ÀÌ´Ù. º» Á¦Ç°Àº genomic DNA purification kit¿Í 200ȸºÐÀÇ TB Green qPCR premix ¹×, calibrated control template¸¦ Æ÷ÇÔÇÏ°í ÀÖ´Ù.

* º» Á¦Ç° ³»¿¡ Æ÷ÇÔµÈ primer´Â Packaging Signal¿¡ ÀÎÁ¢ÇÑ HIV-1 genome ¿µ¿ªÀ» ÁõÆøÇÒ ¼ö ÀÖµµ·Ï µðÀÚÀεǾî ÀÖ½À´Ï´Ù. º» ÇÁ¶óÀ̸Ӵ ´ÙÄ«¶ó¹ÙÀÌ¿À¿¡¼­ ÆǸÅÇÏ´Â ¸ðµç Lentivirus vector »Ó¸¸ ¾Æ´Ï¶ó ÀϹÝÀûÀ¸·Î »ç¿ëµÇ´Â HIV-1 based lentiviral vector¿¡ ȣȯ °¡´ÉÇÕ´Ï´Ù. ¸¸¾à, vector compatibilty¸¦ È®ÀÎÇÏ°íÀÚ ÇÑ´Ù¸é, °í°´Áö¿ø¼¾ÅÍ (TEL. 02-2081-2510)·Î ¿¬¶ô ¹Ù¶ø´Ï´Ù.


±×¸² 1. Measuring lentiviral copy number in target cells. Provirus content is determined using a rapid qPCR method.


±×¸² 2. Lentiviral provirus copy number affects transgene expression levels in transduced cells. HT1080 cells were infected at either high or low MOI using an AcGFP1 lentiviral expression vector. 72 hr after infection, the cells were subjected to puromycin drug selection. One week later, stably transduced clones were selected for expansion and analysis. AcGFP1 expression levels were determined using flow cytometry, and provirus content in genomic DNA was determined using the Lenti-X Provirus Quantitation Kit. The figure represents data pooled from two groups having either low copy number (>3; n = 8) or high copy number (>9; n = 7). Higher provirus contents were associated with higher expression levels.


±×¸² 3. Equivalent virus amounts in different cell types results in different provirus (integrated lentivirus) copy numbers. Equivalent amounts of a single lentiviral stock (100 ¥ìl) were used to infect either HT1080, HeLa, and HEK 293 cells. 72 hr after infection, the cells were subjected to drug selection with puromycin. One week later, the provirus copy numbers of the stably transduced cell populations were determined using the Lenti-X Provirus Quantitation Kit.
±¸¼ºÇ° (ÀÚ¼¼ÇÑ ³»¿ëÀº CoA¸¦ ÂüÁ¶Çϼ¼¿ä)
  • Lenti-X Provirus Quantitation Components
  • NucleoSpin Tissue Kit (10 preps)
  • TB Green Advantage qPCR Premix (200 rxns)
°ü·ÃÁ¦Ç°




Keyword : Lentivirus,virus,Lenti,·»Æ¼,·»Æ¼¹ÙÀÌ·¯½º,Á¤·®,¿ª°¡,titer,titration,provirus,ÇÁ·Î¹ÙÀÌ·¯½º,integrated virus,»ðÀÔ ¹ÙÀÌ·¯½º,moi

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