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Indel (mutation) ¼­¿­ È®ÀÎ

Indel Identification Kit

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
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Clontech
631444
Guide-it¢â Indel Identification Kit
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç°  ¶óÀ̼±½º 
10 ȸ
(Package)
972,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x33613, x32690

* ±¸¼ºÇ° º¯°æ: In-Fusion¢ç HD Cloning Kit (Code 639648)°¡ ¾÷±×·¹À̵å Á¦Ç°ÀÎ In-Fusion¢ç Snap Assembly Master Mix (Code 638947)·Î ´ëüµÇ¾î Á¦°øµË´Ï´Ù.

  • Streamlined method: genome editing ÀÌÈÄ, ´Ù¾çÇÑ Indel sequence È®ÀÎ
  • CRISPR/Cas9 »Ó¸¸ ¾Æ´Ï¶ó TALENs, ZFNs ÀÌÈÄ Indel identification¿¡ »ç¿ë °¡´É
  • Ultrafast protocol: Cell direct PCR°ú In-Fusion CloningÀ» Á¶ÇÕÇÑ ºü¸¥ ÇÁ·ÎÅäÄÝ
  • Complete kit: target ÀÇ ÁõÆø, º¹Á¦, sequencing¿ë »ùÇà Á¦ÀÛ¿¡ ÇÊ¿äÇÑ ¸ðµç ½Ã¾à Æ÷ÇÔ
Á¦Ç°¼³¸í
Guide-it Indel Identification Kit´Â genome editing (CRISPR/Cas9, TALENs, ZFNs) ÀÌÈÄ ¾ò¾îÁøIndel (Insertions and deletions)ÀÇ ´Ù¾ç¼ºÀ» È®ÀÎÇÏ´Â µ¥ ÃÖÀûÈ­µÈ Á¦Ç°ÀÌ´Ù. º» Á¦Ç°Àº DNA ¼­¿­ ºÐ¼®À» À§ÇÑ target siteÀÇ amplify, clone, sequencing¿ë »ùÇà Á¶Á¦¸¦ À§ÇÑ ¸ðµç Á¦Ç°°ú ÇÁ·ÎÅäÄÝÀ» ±¸ºñÇÏ°í ÀÖ¾î ½Å¼ÓÇÏ°í ½Å·Ú¼º ³ôÀº °á°ú¸¦ ¾òÀ» ¼ö ÀÖ´Ù. Terra PCR Direct polymerase¸¦ »ç¿ëÇϹǷΠDNA ÃßÃâ, Á¤Á¦ ¾øÀÌ crude cell lysate·ÎºÎÅÍ genomic DNA fragment¸¦ ÁõÆøÇÒ ¼ö ÀÖÀ¸¸ç, ÁõÆø»ê¹°Àº In-Fusion Cloning ¹æ¹ýÀ» ÀÌ¿ëÇÏ¿© pre-linearized pUC19¿¡ ¹Ù·Î cloning ÇÒ ¼ö ÀÖ´Ù. ÀÌÈÄ Á¦Ç° ³»¿¡ Æ÷ÇÔµÈ Stellar Competent Cell¿¡ transformation ÀÌÈÄ »ý¼ºµÈ Colony´Â colony PCR ÇÏ¿© target regionÀ» ÁõÆøÇÑ ÈÄ ½ÃÄö½ÌÇÏ¿© Indel ¼­¿­À» ºÐ¼®ÇÒ ¼ö ÀÖ´Ù (±×¸² 1, ±×¸² 2).



±×¸² 1. The Guide-it Indel Identification Kit provides a complete workflow for identifying the variety of insertions and deletions (indels) introduced by nuclease-based genome editing. The protocol uses direct PCR to amplify a genomic DNA fragment (~500 to 700 bp) containing the target site directly from crude cell lysates (step 1). The resulting amplified fragments contain a pool of edited target sites from individual cells. These PCR products are cloned directly into a pre-linearized pUC19 vector using the In-Fusion cloning system (step 2). After transformation of an optimized E. coli strain, colony PCR is used to amplify the target site from the plasmid (step 3). DNA sequencing is then used to identify the different indels generated at the targeted genomic site (step 4).



±×¸² 2. Identification of insertions and deletions (indels) in the CD81 gene after CRISPR/Cas9 targeting. HeLa cells were transfected with plasmids encoding Cas9 and an sgRNA targeting the CD81 gene. The Guide-it Indel Identification Kit was used to prepare CD81 target sites for DNA sequence analysis. The sequencing data from six clones was aligned with the wild-type sequence, revealing a broad range of indels in the CD81 gene.
Àû¿ë
  • Detecting insertions and deletions introduced in mammalian genomic DNA
  • Characterizing the variety of indels that are present in a cell population after nuclease-based gene editing
±¸¼ºÇ° (ÀÚ¼¼ÇÑ ³»¿ëÀº CoA¸¦ ÂüÁ¶Çϼ¼¿ä)
  • Guide-it Indel Identification Components
    • 110 ¥ìl Terra PCR Direct Polymerase Mix
    • 3 x 1 ml 2X Terra PCR Direct Buffer (with Mg2+, dNTP)
    • 400 ¥ìl Extraction Buffer 1
    • 40 ¥ìl Extraction Buffer 2
    • 10 ¥ìl pUC19 Cloning Vector, linearized (50 ng/¥ìl)
    • 200 ¥ìl Colony PCR Forward Primer (15 ¥ìM)
    • 200 ¥ìl Colony PCR Reverse Primer (15 ¥ìM)
    • 6 x 1 ml PCR-Grade Water
  • In-Fusion Snap Assembly Master Mix
  • NucleoSpin Gel and PCR Clean-Up Kit
  • Stellar Competent Cells
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Guide-it Indel Identification

-20¡É

In-Fusion Snap Assembly Master Mix

-20¡É

NucleoSpin Gel and PCR Clean-up

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Stellar Competent Cells:-70¡É

-70¡É

±× ¿Ü ±¸¼ºÇ°

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Keyword : Indel,mutation È®ÀÎ,Insertion,Deletion,NHEJ,Knockout,Knock-out,Knock out,KO,CRISPR,CRISPR/Cas9,Cas9,sgRNA,Å©¸®½ºÆÛ,gene editing,À¯ÀüÀÚÆíÁý

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