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sgRNA ÇÕ¼º°ú ½ºÅ©¸®´×

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
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Clontech
632635
Guide-it¢â sgRNA In Vitro Transcription Kit
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
50ȸ
(Package)
1,445,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x101786
Clontech
632636
Guide-it¢â Complete sgRNA Screening System
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
50ȸ
(Package)
1,776,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x101786
Clontech
632639
Guide-it¢â sgRNA Screening Kit
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
50ȸ
530,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x101786
Clontech
632638
Guide-it¢â IVT RNA Clean-Up Kit
°ü·ÃÇмú ±¸¸ÅÇϱâ
50ȸ
741,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x101786

  • CRISPR/Cas9 genome editingÀ» À§ÇÑ sgRNAÀÇ ÇÕ¼º (in vitro transcription)
  • °£ÆíÇÑ ÇÁ·ÎÅäÄÝ·Î °íÇ°Áú, °í¼öÀ² (12ug ÀÌ»ó)ÀÇ sgRNA Á¦ÀÛ °¡´É
  • ÇÕ¼ºÇÑ sgRNA´Â ÃßÈÄ delivery (transfection, electroporation)¿¡ ÀÌ¿ë °¡´É
  • Screening Kit¿¡ Æ÷ÇԵǾîÀÖ´Â Cas9 ´Ü¹éÁú°ú Á¶ÇÕÇÏ¿©, ½ÇÁ¦ ¼¼Æ÷ ½ÇÇè Àü¿¡ sgRNAÀÇ À¯È¿¼º ¹× È¿À² È®ÀÎ °¡´É (sgRNA screening)
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Guide-it¢â sgRNA In vitro Transcription Kit (Code 632635)´Â in vitro transcription ¹ÝÀÀÀ¸·Î °£ÆíÇÏ°Ô 12ug ÀÌ»óÀÇ °íÇ°Áú, °í¼öÀ²ÀÇ sgRNA¸¦ Á¦ÀÛÇÒ ¼ö ÀÖ´Ù. º» Á¦Ç° ³»¿¡´Â Phenol/Chloroform ¹æ½ÄÀÌ ¾Æ´Ñ ½ºÇÉ Ä÷³ ¹æ½ÄÀÇ RNA Á¤Á¦ Å°Æ®ÀÎ Guide-it¢â IVT RNA Clean-Up Kit (Code 632638)°¡ Æ÷ÇԵǾîÀÖ´Ù. Á¤Á¦ÇÑ sgRNA´Â ±×´ë·Î transfection, electroporation ¶Ç´Â in vitro cleavage assay¿¡ Àû¿ëÇÒ ¼ö ÀÖ´Ù.
Guide-it¢â Complete sgRNA Screening System (Code 632636)Àº Guide-it¢â sgRNA In Vitro Transcription Kit (Code 632635)¿Í Guide-it¢â sgRNA Screening Kit (Code 632639) Á¦Ç°À¸·Î ±¸¼ºµÇ¾îÀÖ´Ù. º» Á¦Ç°À» »ç¿ëÇÏ¿© in vitro transcriptionÀ¸·Î ÇÕ¼ºÇÑ sgRNA¿Í Á¦Ç° ³» Æ÷ÇÔµÈ Cas9 ÀçÁ¶ÇÕ ´Ü¹éÁúÀ» ½ÇÇè Æ©ºê ³»¿¡¼­ ¹ÝÀÀ½ÃÅ´À¸·Î½á, ½ÇÁ¦ ¼¼Æ÷ ½ÇÇè Àü¿¡ in vitro cleave assay¸¦ ÅëÇÏ¿© sgRNAÀÇ À¯È¿¼º ¹× Àý´Ü È¿À²À» È®ÀÎ °¡´ÉÇÏ´Ù. º» Á¦Ç° ÀÌ¿ë ½Ã, sgRNAÀÇ Àý´ÜºÎÀ§¸¦ Æ÷ÇÔÇÑ DNA´Â PCR ÁõÆø ÈÄ, ÀçÁ¶ÇÕ Cas9 nuclease·Î Àý´ÜÀ» Çϸç, ÀÌ Àý´Ü È¿À²Àº agarose gel eletrophoresis·Î È®ÀÎÇÒ ¼ö ÀÖ´Ù.
±âÁ¸ Á¦Ç°°úÀÇ Â÷ÀÌÁ¡
º» Á¦Ç°Àº ±âÁ¸ Á¦Ç°°ú Á¦Ç°¸íÀº °°Áö¸¸, ¹öÀü ¾÷±×·¹À̵å Ãâ½ÃµÇ¾úÀ¸¸ç ¾Æ·¡¿Í °°ÀÌ º¯°æµÇ¾ú´Ù.

Guide-it¢â sgRNA In Vitro Transcription Kit
  • ¿ë·®: 10ȸ ¡æ 50ȸ·Î º¯°æ
  • ÇÁ·Î¸ðÅÍ ¹× scaffold ¼­¿­ º¯ÇüÀ¸·Î sgRNA ÇÕ¼º ¼öÀ² 3¹è ÀÌ»ó Çâ»ó(12ug ÀÌ»ó) ¹× genome editing È¿À² Çâ»ó
  • Forward primer ¼­¿­ º¯°æ: T7 promoter¿Í scaffold ¼­¿­ º¯ÇüÀ¸·Î Primer ÃÑ ±æÀÌ°¡ 69nt ¡æ 56-58nt·Î º¯°æ

[±âÁ¸]- total length 69nt


[½Å±Ô]- total length 56-58nt


  • Template DNA(sgRNA encoding template)ÀÇ PCR ÁõÆøÀ» À§ÇÏ¿© PrimeSTAR¢ç Max DNA Polymerase¸¦ Àû¿ëÇÏ¿© ºñƯÀÌÀû ÁõÆø °¨¼Ò, Agarose gel purification ¹æ¹ý ºÒÇÊ¿ä
  • Phenol/Chloroform Á¤Á¦ ´ë½Å ½ºÇÉ Ä÷³ ¹æ½ÄÀÇ RNA Clean-Up KitÀ¸·Î RNA Á¤Á¦

Guide-it¢â sgRNA Screening Kit
  • ¿ë·®: 30ȸ ¡æ 50ȸ·Î º¯°æ
  • Target DNA(DNA cleavage template)ÀÇ Á¶Á¦¿¡ Terra PCR MixÀ» ÀÌ¿ëÇÏ¿© Direct PCR °¡´É
»ó¼¼³»¿ë ¹× µ¥ÀÌÅÍ

±×¸² 1. Flow chart describing how the Guide-it sgRNA In Vitro Transcription Kit (including the Guide-it IVT RNA Clean-Up Kit) and Guide-it sgRNA Screening Kit work together in the Guide-it Complete sgRNA Screening System, which can be used to synthesize and test the efficacy of sgRNAs.


±×¸² 2. Schematic highlighting the key steps for using Guide-it kits for the production of high amounts of single guide RNA (sgRNA) and screening for cleavage efficacy at the desired target site using recombinant Cas9 protein.


±×¸² 3. RNA quality produced via in vitro transcription. sgRNAs produced using the Guide-it sgRNA In Vitro Transcription Kit were compared with sgRNAs produced using a competitor¡¯s kit. The Guide-it kit produced a high-quality, single band for each reaction, while the competitor¡¯s kit showed unwanted byproducts.


±×¸² 4. sgRNA yield and quality for different gene targets using the Guide-it Complete sgRNA Screening System. Panel A. 20 ¥ìl of each sgRNA in vitro transcription (IVT) reaction was incubated for 4 hr at 37¡ÆC. The yield for each transcribed sgRNA was quantified using a NanoDrop spectrophotometer. Panel B. Each sgRNA IVT reaction was a run on an Agilent Bioanalyzer to assay for sample quality.
º¸Á¸
Guide-it¢â sgRNA In Vitro Transcription Components v2: -20¡É
Guide-it¢â IVT RNA Clean-Up Kit: ½Ç¿Â
Guide-it¢â sgRNA Screening Kit: -20¡É
±¸¼ºÇ° (ÀÚ¼¼ÇÑ ³»¿ëÀº CoA¸¦ ÂüÁ¶Çϼ¼¿ä)
Guide-it¢â sgRNA In Vitro Transcription Kit (Code 632635; 50ȸ)
  • 1 each Guide-it sgRNA In Vitro Transcription Components v2
    • 50 ¥ìl Guide-it Scaffold Template (1 ng/¥ìl)
    • 350 ¥ìl Guide-it In Vitro Transcription Buffer
    • 1 ml RNase Free Water
    • 500 ¥ìl RNase Free Water
    • 150 ¥ìl Guide-it T7 Polymerase Mix (33 U/¥ìl)
    • 625 ¥ìl PrimeSTAR Max Premix (2X)
    • 100 ¥ìl Recombinant DNase I (RNase-Free) (5 U/¥ìl)
  • 1 each Guide-it IVT RNA Clean-Up Kit (Code. 632638; 50 rxns)
    • 13 ml RNase Free Water
    • 3 ml IVT Binding Buffer
    • 50 IVT RNA Clean-Up Spin Columns
    • 2 x 6 ml IVT Wash Buffer
    • 50 Collection Tubes (2 ml)

Guide-it¢â IVT RNA Clean-Up Kit(Code 632638; 50ȸ)
  • 13 ml RNase Free Water
  • 3 ml IVT Binding Buffer
  • 50 IVT RNA Clean-Up Spin Columns
  • 2 x 6 ml IVT Wash Buffer
  • 50 Collection Tubes (2 ml)

Guide-it¢â sgRNA Screening Kit (Code 632639; 50ȸ)
  • 25 ¥ìl Guide-it Recombinant Cas9 Nuclease (500 ng/¥ìl)
  • 50 ¥ìl 15X Cas9 Reaction Buffer
  • 50 ¥ìl 15X BSA
  • 5 ¥ìl Control sgRNA (50 ng/¥ìl)
  • 25 ¥ìl Control Fragment (20 ng/¥ìl)
  • 50 ¥ìl Terra PCR Direct Polymerase Mix (1.25 U/¥ìl)
  • 1.25 ml 2X Terra PCR Direct Buffer (with Mg2+, dNTP)
  • 5 ml RNase Free Water
  • 5 ml Extraction Buffer 1
  • 0.5 ml Extraction Buffer 2

Guide-it¢â Complete sgRNA Screening System (Code 632636) Á¦Ç°Àº Guide-it¢â sgRNA In Vitro Transcription Kit (Code 632635)¿Í Guide-it¢â sgRNA Screening Kit (Code 632639)¸¦ Æ÷ÇÔÇÏ°í ÀÖ½À´Ï´Ù.

Keyword : CRISPR,CRISPR/Cas9,Cas9,sgRNA,RNA prep,Guide-it,genome editing,Å©¸®½ºÆÛ, 631439, 631438, 631440, RNP, Ribonucleoprotein, Ribonucleo-protein