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Baculovirus ¿ª°¡£¨Genomic DNA copy ¼ö£©¸¦ ¾à 4 ½Ã°£¿¡ ÃøÁ¤

qPCR Titer ÃøÁ¤ Kit

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Clontech
631414
BacPAK qPCR Titration Kit
°ü·ÃÇмú ±¸¸ÅÇϱâ
200 ȸ
(Package)
978,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x102919



  • ºü¸¥ titration kit¸¦ ÀÌ¿ëÇÏ¿© 4 ½Ã°£ ¸¸¿¡ ¹ÙÀÌ·¯½º ¿ª°¡¸¦ ÃøÁ¤
  • ÇÏ·ç ¾È¿¡ virusÀÇ ¿ª°¡ ÃøÁ¤ ¹× °¨¿°±îÁö °¡´É
  • AcMNPV ±â¹ÝÀÇ ¸ðµç baculovirus¿¡µµ ÀûÇÕ
BacPAK qPCR Titration Kit´Â ¹ÙÀÌ·¯½ºÀÇ ¿ª°¡ ÃøÁ¤À» À§ÇÑ ¸Å¿ì ºü¸£°í °£´ÜÇÑ ¹æ¹ýÀ» Á¦°øÇÑ´Ù. ÀÌ kit´Â Real Time PCR°ú Intercalater ±â¼úÀ» »ç¿ëÇÏ¿© viral genomeÀ» ±¸¼ºÇϱâ Àü¿¡ ºü¸£°Ô DNA Á¤Á¦ ´Ü°è¸¦ ÁøÇàÇÑ´Ù (Figure 1). Ç¥ÁØ ¿ª°¡ ÃøÁ¤ ¹æ¹ýÀº ÃøÁ¤À» ¿Ï·áÇϱâ±îÁö ÃÖ´ë 10 ÀÏÀÌ ¼Ò¿äµÇ´Â ¹Ý¸é¿¡, º» Á¦Ç°À» ÀÌ¿ëÇϸé 4 ½Ã°£¸¸¿¡ ¿ª°¡ÃøÁ¤ÀÌ ¿Ï·áµÇ¸ç AcMNPV ±â¹ÝÀÇ ¾î¶² baculoviral vector¿¡µµ »ç¿ë °¡´ÉÇÏ´Ù. Real Time PCR¹ýÀ» ÀÌ¿ëÇÏ¿© ¹ÙÀÌ·¯½º ³óÃà°ú ¸ñÀû ¼¼Æ÷ °¨¿° »çÀÌÀÇ ½Ã°£ °£°ÝÀ» Å©°Ô ÁÙ¿© ´çÀÏ¿¡ ¸ðµÎ ¼öÇàÇÒ ¼ö ÀÖµµ·Ï ÇØÁØ´Ù. ¹ÙÀÌ·¯½ºÀÇ °¨¿°´É·ÂÀÌ ÀúÇϵǴ °ÍÀ» ÇÇÇÒ ¼ö ÀÖ°í, Á¤È®ÇÑ multiplicity of infection (MOI)À¸·Î ¸ñÀû ¼¼Æ÷¸¦ °¨¿°½ÃÄÑ ´õ ÀÏ°üµÈ °á°ú¸¦ µµÃâÇÒ ¼ö ÀÖ´Ù (1).

BacPAK qPCR Titration Kit´Â Á¶Á¤µÈ DNA standard curve·ÎºÎÅÍ baculovirus Áغñ ´Ü°è¿¡ ÇÊ¿äÇÑ viral genome copy¼ö¸¦ ÃøÁ¤Çϵµ·Ï ÇÑ´Ù (Figure2). ÀýÂ÷´Â °£´ÜÇÏ¿© viral DNA¿Í BacPAK control DNA´Â ´Ü°èÀûÀ¸·Î Èñ¼®(serial dilution)ÇÏ¿© qPCRÀ» ÁغñÇÏ°í, °¢ ¹ÙÀÌ·¯½º ½Ã·áÀÇ DNA copy ¼ö´Â Figure 2¿¡ ³ªÅ¸³­ ´ë·Î °¢°¢ÀÇ copy ¼ö¿¡ ´ëÇÑ Èñ¼®µÈ control ½Ã·áÀÇ Ct °ªÀ» °î¼±È­ÇÏ¿© »ý¼ºµÇ´Â standard curve¿Í Ct °ªÀ» ºñ±³ÇÏ¿© ÃøÁ¤µÈ´Ù. °£´ÜÇÏ°í, ÀçÇö¼ºÀÌ ³ôÀ¸¸ç, ªÀº ÃøÁ¤ ½Ã°£À¸·Î BacPAK qPCR Titration Kit´Â baculoviral DNA titer ÃøÁ¤¿¡ ¾ÆÁÖ ÀûÇÕÇÏ´Ù.


Figure 1. The BacPAK qPCR Titration Kit protocol.


Figure 2. The BacPAK qPCR Titration Kit exhibits a wide dynamic range. The BacPAK DNA Control Template was serially diluted from 108 to 103 copies per sample and analyzed with the BacPAK qPCR Titration Kit. The amplification plots (Panel A) show a wide dynamic range of at least 6 orders of magnitude with no NTC (No-Template Control) background. The standard curve (Panel B) obtained by plotting the Ct values (determined from the amplification plots in Panel A) against the log of the DNA copy number in each sample, demonstrates a strong linear correlation between the Ct and the DNA copy number (log scale), with R2 = 1.000 and a PCR efficiency of 100%.


* Clontech offers two different kits for baculovirus titration: the BacPAK Baculovirus Rapid Titer Kit (Cat. No. 631406) utilizes a standard immunological assay to accurately identify virus-infected cells, and the BacPAK qPCR Titration Kit (Cat. No. 631414) measures viral DNA copies via SYBR qPCR.
Components & Storage Conditions
°¢ Á¦Ç° ±¸¼º¹°°ú º¸°üÁ¶°ÇÀº Certificate of Analysis ¸¦ ÂüÁ¶ÇϽʽÿÀ.
References
1. Rapid & Accurate Baculovirus Titration (2009) Clontechniques XXIV(3):8-9.

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