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Lentivirus³ª MMLV retrovirusÀÇ ´õ ºü¸¥ transduction À¯µµ

Transduction ½Ã¾à

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
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Clontech
631254
Lenti-X¢â Accelerator Starter Kit
°ü·ÃÇмú ±¸¸ÅÇϱâ
each
(Package)
2,089,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x32762
Clontech
631256
Lenti-X¢â Accelerator
°ü·ÃÇмú ±¸¸ÅÇϱâ
400 ¥ìl
676,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x32762
Clontech
631257
Lenti-X¢â Accelerator
°ü·ÃÇмú ±¸¸ÅÇϱâ
1,000 ¥ìl
1,662,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Clontech
631255
Magnetic Separator for Cell Culture
°ü·ÃÇмú ±¸¸ÅÇϱâ
each
1,553,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â


Polybrene¾øÀÌ 25ºÐ³» lentiviral transductionÀ» ÃËÁøÇÑ´Ù.

- Lentivirus³ª MMLV retrovirusÀÇ ´õ ºü¸¥ transduction À¯µµ
- Stem cell°ú °°ÀÌ ¹Î°¨ÇÑ ¼¼Æ÷¿¡ ÀÌ»óÀû
- Starter KitÀº magnetic separator¸¦ Æ÷ÇÔÇÑ´Ù

Lenti-X Accelerator´Â lentivirus¿Í MSCV retrovirus¸¦ Æ÷ÇÔÇÑ MMLV retrovirusÀÇ transductionÀ» ÃËÁøÇϵµ·Ï µðÀÚÀÎµÈ magnetic bead ±â¹ÝÀÇ ±â¼úÀÌ´Ù. Lenti-X Accelerator´Â beadµé virus¿Í °áÇÕ½ÃÅ°±â À§ÇØ 20ºÐµ¿¾È preincubationÇÑ µÚ, ´ÜÁö 5ºÐ µ¿¾È¸¸ ¼¼Æ÷¸¦ ¹ÙÀÌ·¯½º »óÃþ¾×(viral supernatant)¿¡ ³ëÃâ½ÃÅ°±â ¶§¹®¿¡ ¸Å¿ì ¹Î°¨ÇÑ ¼¼Æ÷ÀÇ Àû¿ë¿¡ ÀÌ»óÀûÀÌ´Ù. ¹Ý¸é ¼¼Æ÷¿¡ polybreneÀ» ó¸®Çϸé ÇÏ·ç¹ãµ¿¾È incubationÀ» ÇؾßÇÑ´Ù.

±×¸² 1

Figure 1. Transduction with Lenti-X Accelerator. Placing virus-bound magnetic beads in a magnetic field greatly increases the localized virus concentration at the surface of the cell monolayer. This reduces the transduction time to just 5 min after a 20 min preincubation to bind the beads to the virus-compared to transduction overnight with Polybrene. Accelerated transduction also limits exposure of your sensitive target cells to viral supernantant.
±×¸² 2

Figure 2. Lenti-X Accelerator provides high transduction efficiency in a 25 min protocol. A ZsGreen1-expessing Lenti-X vector was used to transduce HT1080 cells. Lenti-X Accelerator beads (8 §¡) were preincubated for 20 min at room temperature with 200 §¡ (approx. 1x106 total IFU) of viral supernatant, and applied to HT1080 cells on a Magnetic Separator for 5 min. HT1080 cells were also transduced with the same amount of virus in the presence of 6 §¶/ml Polybrene, for 5 min or overnight. After the cultures were grown for an additional 72 hr at 37¡É, the number of transduced cells was determined by flow cytometry.




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