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Home > ÀüÁ¦Ç°º¸±â > Genome Editing > À¯ÀüÀÚ ÆíÁý Àü ´Ü°è > [Upgrade] Guide-it¢â Long ssDNA Production System v2
À¯ÀüÀÚ »ðÀÔÀ» À§ÇÑ donor template ÇÕ¼º (5 kb ssDNA)

[Upgrade] Guide-it¢â Long ssDNA Production System v2

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Clontech
632666
Guide-it¢â Long ssDNA Production System v2
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(Package)
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* º» Á¦Ç°Àº ±âÁ¸ Guide-it¢â Long ssDNA Production System (Code 632644)ÀÇ ¾÷±×·¹ÀÌµå ¹öÀü Á¦Ç°ÀÔ´Ï´Ù.
¼º°øÀûÀÎ Knock-inÀ» À§ÇÑ FAQ È®ÀÎ (Ŭ¸¯)

  • ´Ù¾çÇÑ gene editing toolÀÇ À¯ÀüÀÚ »ðÀÔÀ» À§ÇÑ donor template ÇÕ¼º
  • 500bp ~ 5kb ±îÁöÀÇ single strand DNA (ssDNA) ÇÕ¼º °¡´É
  • Cloning°ú Gel selection Á¤Á¦ °úÁ¤ÀÌ ¾ø´Â ºü¸£°í ½¬¿î ÇÁ·ÎÅäÄÝ
  • Off target ÃÖ¼ÒÈ­ - dsDNA donor template¿¡ ºñÇÏ¿© ±Øµµ·Î ³·Àº randomÇÑ off-target integration·Î ³ôÀº Editing È¿À²
  • ¼¼Æ÷µ¶¼ºÀ» ÃÖ¼ÒÈ­ÇÏ¿©, ¼¼Æ÷ »ýÁ¸À² Çâ»ó
Á¦Ç°¼³¸í
À¯ÀüÀÚ »ðÀÔÀ» À§ÇÑ Donor repair templateÀÇ °æ¿ì double-stranded DNA (dsDNA)¿¡ ºñÇÏ¿© single-stranded DNA (ssDNA) repair template°¡ º¸´Ù ¸¹Àº ÀÌÁ¡ÀÌ ÀÖ´Ù°í ¾Ë·ÁÁ® ÀÖÁö¸¸, long ssDNA donor templateÀÇ ÇÕ¼º°ú ºñ¿ëÀÇ ¹®Á¦°¡ ÀÖ¾ú´Ù.
Guide-it¢â Long ssDNA Production System v2´Â ´Ù¾çÇÑ gene editing toolÀ» ÀÌ¿ëÇÑ knock-in ½ÇÇèÀ» À§ÇÑ ÃÖÀå 5kbÀÇ ssDNA repair template oligonucleotide¸¦ 50ȸ ÇÕ¼ºÇÒ ¼ö ÀÖ´Â Á¦Ç°ÀÌ´Ù. ±âÁ¸ Á¦Ç°¿¡ ºñÇØ StrandaseÀÇ ¼º´ÉÀÌ ¾÷±×·¹ÀÌµå µÇ¾úÀ» »Ó ¾Æ´Ï¶ó, ÃÖÀûÀÇ buffer¿Í Á¤Á¦Á¦Ç°±îÁö Æ÷ÇÔÇÏ°í ÀÖ´Ù. º» Á¦Ç°À» ÀÌ¿ëÇϸé Strandase¸¦ ó¸®ÇÏ´Â °Í¸¸À¸·Îµµ dsDNA PCR »ê¹°À» sense ¶Ç´Â antisense strand·Î ºÐ¸®ÇÏ¿© ssDNA¸¦ Á¦ÀÛÇÒ ¼ö ÀÖ´Ù.

ssDNA donor templateÀÇ ÀåÁ¡


±×¸² 1. Schematic describing the steps involved in the preparation of long ssDNA donors for use in HDR experiments.


±×¸² 2. Guide-it Long ssDNA Production System v2¸¦ ÀÌ¿ëÇØ °³¼±µÈ ssDNA ÇÕ¼º ¼º´É
dsDNA¿Í ±âÁ¸ Á¦Ç°À¸·Î ÇÕ¼ºÇÑ ssDNA¿Í Àü±â¿µµ¿À¸·Î ºÐ¼®ÇßÀ» ¶§, ¾÷±×·¹À̵åµÈ Guide-it Long ssDNA Production System v2·Î ÇÕ¼ºÇÑ ssDNA¿¡¼­ °¡Àå ¼±¸íÇÑ ¹êµå·Î È®ÀεǾú´Ù.
Gel image showing the dsDNA starting material (Lane 1) and sense (SS) or antisense (AS) ssDNA products for several HDR templates generated using the original (v1) or updated (v2) versions of the Guide-it Long ssDNA Production System (Lane 2 and Lane 3, respectively). dsDNA templates included in this analysis had been identified as challenging substrates for ssDNA generation using the v1 Guide-it kit. The dsDNA and ssDNA were analyzed via agarose gel electrophoresis using ethidium bromide as a staining agent. ssDNA products run at a smaller molecular weight than corresponding dsDNA substrates. In all cases, the Guide-it Long ssDNA Production System v2 generated a cleaner band of ssDNA than the previous version of the kit, suggesting more complete and uniform digestion.
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Keyword : Knock-in,KI,Knock in,Knockin,ssDNA,HDR template,donor template,repair template,HDR pathway,CRISPR,CRISPR/Cas9,Cas9,sgRNA,Å©¸®½ºÆÛ,gene editing,À¯ÀüÀÚÆíÁý

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