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Gene editing
> [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
50ȸ
(Package)
1,257,000¿ø
Product index
Overview
RetroNectin¢ç - Viral ..
[FAQ] Retronectin
RetroNectin¢çÀÇ ´Ù¾çÇÑ..
T cell ¿¬±¸ÀÚ¸¦ À§ÇÑ ..
[Cancer Research] Immu..
Cancer ¿¬±¸ °¡À̵å
T cell therapy ¿¬±¸
RetroNectin¢ç (Recombi..
Anti-CD3 mAb GMP grade
LymphoONE¢ç T-Cell Exp..
Cytokine GoStix¢â Plus..
GT-T551 Culture medium
¼¼Æ÷ ¹è¾ç¿ë °¡½º Åõ°ú..
Gene editing
Recombinant Cas9 ´Ü¹é..
sgRNA ÇÕ¼º°ú ½ºÅ©¸®´×
TransIT-X2¢ç Transfect..
[Upgrade] Guide-it¢â L..
TCR/BCR Profiling
Human TCR a/b Profilin..
Mouse TCR a/b Profilin..
Mouse BCR IgG H/K/L Pr..
¼¼Æ÷¹è¾ç °ü·Ã Á¦Ç°
Cell culture ÇʼöÇ° - ..
¼¼Æ÷¹è¾ç½Ã Mycoplasma ..
MycoAlert PLUS mycopla..
MycoAlert Mycoplasma D..
MycoZap Antibiotics
MycoZap Mycoplasma Eli..
TaKaRa PCR Mycoplasma ..
CycleavePCR Mycoplasma..
Ç×»ýÁ¦ ½Ã¸®Áî I
Phosphate Buffered Sal..
TBS(Tris-Buffered Sali..
* º» Á¦Ç°Àº ±âÁ¸ Guide-it¢â Long ssDNA Production System (Code 632644)ÀÇ ¾÷±×·¹ÀÌµå ¹öÀü Á¦Ç°ÀÔ´Ï´Ù.
´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
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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¡
Significantly reduced random and off-target integration
, resulting in improved gene editing efficiency and lower likelihood of experimental artefacts
Minimal cytotoxicity
, enabling recovery of larger populations of edited cells
Negligible transgene expression from nonintegrated templates, allowing for easier identification of correctly edited clones
±×¸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 :
Gene editing
Recombinant Cas9 ´Ü¹éÁú
sgRNA ÇÕ¼º°ú ½ºÅ©¸®´×
Trans
IT-X2¢ç Transfection Reagent
À¯ÀüÀÚ »ðÀÔÀ» À§ÇÑ donor template ÇÕ¼º (5 kb..
[Upgrade] Guide-it¢â Long ssDNA Production System v2