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Home > ÀüÁ¦Ç°º¸±â > PCR > Mutagenesis > Diversify PCR Random Mutagenesis Kit
Error prone PCR¿¡ ÃÖÀû

Diversify PCR Random Mutagenesis Kit

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Clontech
630703
Diversify PCR Random Mutagenesis Kit
°ü·ÃÇмú ±¸¸ÅÇϱâ
30 ȸ
1,580,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x32608, x32607

  • Ÿ°Ù DNA ¼­¿­ÀÇ mutation rateÀ» Á¤¹ÐÇÏ°Ô Á¶Àý
  • Hot spot ¾øÀÌ ±¤¹üÀ§ÇÑ mutation À¯µµ °¡´É
  • ¸ðµç Á¾·ùÀÇ ¿°±â¼­¿­ ġȯ °¡´É
  • Random mutationÀ» Á¶ÀýÇÏ¿© ´Ü¹éÁú ±â´É ¿¬±¸¿¡µµ À¯¿ë
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Diversify™ PCR Random Mutagenesis Kit´Â µ¶¼ºÀÌ ¾ø´Â ¹æ¹ýÀ» »ç¿ëÇÏ¿© ÃÖ´ë 4 kb ±æÀÌÀÇ ¼­¿­¿¡ random mutationÀ» À¯¹ßÇϱâ À§ÇÑ Á¦Ç°ÀÌ´Ù.

- Diversify ¹æ¹ý
º» Á¦Ç°À» »ç¿ëÇϸé mutagenesis À¯¹ß Á¶°ÇÀ» ½±°Ô Á¶ÀýÇÒ ¼ö À־ ¸Å¿ì ´Ù¾çÇÑ »çÀÌÁîÀÇ DNA ¼­¿­¿¡µµ ÃÖÀûÀÇ mutation¸¦ µµÀÔÇÒ ¼ö ÀÖ´Ù.
Mn2+¿Í dGTPÀÇ ¾çÀ» º¯È­½ÃÅ°´Â °ÍÀ¸·Î mutation rateÀ» 1,000 bp´ç 2~8°³±îÁö ¼±ÅÃÇÏ´Â °ÍÀÌ °¡´ÉÇÏ´Ù.

Diversify ¹æ¹ý¿¡¼­´Â ClontechÀÇ Titanium Taq PCR È¿¼Ò ½Ã½ºÅÛÀ» ÀÌ¿ëÇÏ¿© 4 kb±îÁö DNA ´ÜÆíÀ» ¾ÈÁ¤ÀûÀ¸·Î ÁõÆøÇÑ´Ù. ÀÌ¿Í °°ÀÌ ´Ù¾çÇÑ »çÀÌÁîÀÇ DNA ¼­¿­À» ÁõÆøÇÒ ¼ö Àֱ⠶§¹®¿¡ ªÀº DNA ´ÜÆíÀº ¹°·Ð, operon, plasmid ±×¸®°í Å©±â°¡ Å« ´Ü¹éÁúÀ» ÄÚµùÇÏ´Â ¼­¿­¿¡ mutationÀ» µµÀÔÇϱ⿡µµ ÀÌ»óÀûÀÌ´Ù.

Diversify™ PCR Random Mutagenesis Kit´Â TITANIUM® Taq DNA Polymerase (Code 639208)¿Í ÇÔ²² »ç¿ëÇϵµ·Ï ÃÖÀûÈ­µÇ¾î ÀÖÀ¸¸ç, TaqStart™ Antibody (Code 639250)¸¦ Æ÷ÇÔÇÏ´Â Titanium Taq PCR È¿¼Ò ½Ã½ºÅÛ°ú mutationÀ» À¯¹ßÇÏ´Â buffer¿Í ½Ã¾àÀÌ ¸ðµÎ Æ÷ÇԵǾî ÀÖ´Ù.


±×¸² 1.  Buffer Á¶¼º¿¡ µû¸¥ mutation rate Á¶Àý. ´Ù¸¥ dNTPsÀÇ ³óµµ´Â °íÁ¤ÇÑ Ã¤ Mn2+¿Í dGTPÀÇ ³óµµ¸¦ º¯È­½ÃÅ´À¸·Î½á mutagenesis Á¤µµ¸¦ Á¶ÀýÇÒ ¼ö ÀÖ´Ù. Buffer 1, 5, 9¹ø¿¡¼­ mutation rate´Â DNA sequencingÀ» ÅëÇØ Á÷Á¢ ÃøÁ¤ÇÏ¿´´Ù (°¢ >15,000 bp). ´Ù¸¥ mutations rateÀº Á¤·®Àû in vivo fidelity assayÀÇ mutagenesis level°ú DNA ¿°±â¼­¿­ ºÐ¼® °á°ú¸¦ ¿¬°ü½ÃÄÑ °áÁ¤ÇÏ¿´´Ù.


±×¸² 2. Diversify™ PCR Random Mutagenesis Kit¿¡ ÀÇÇÑ 4 kb ´ÜÆíÀÇ ÁõÆø. ±×¸² 1ÀÇ buffer Á¶°Ç¿¡¼­ 4 kb plasmid (lane 1-9)¸¦ ÁõÆøÇÑ °á°ú¸¦ Àü±â¿µµ¿À» ÅëÇØ È®ÀÎÇÏ¿´´Ù. Å« »çÀÌÁîÀÇ ¿°±â¼­¿­ÀÌ Àß ÁõÆøµÈ °ÍÀ» È®ÀÎÇÏ¿´À¸¸ç, ÀÌ·ÐÀûÀ¸·Î ´õ ±ä ´ÜÆíÀÇ DNA ¿ª½Ã mutagenesis ¹× ÁõÆøÀÌ °¡´ÉÇÏ´Ù.
³»¿ë (30 PCR rxns)
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50X Titanium Taq DNA Polymerase 30 §¡
10X Titanium Taq PCR Buffer 200 §¡
50X Diversify dNTP Mix 30 §¡
50X dNTP Mix (10 mM each) 40 §¡
dGTP (2 mM) 150 §¡
Manganese Sulfate (MnSO4 ; 8 mM) 120 §¡
Control PCR Template (~1ng/§¡) 10 §¡
Control Primer Mix (10 µM each) 10 §¡
Taq I Restriction Enzyme (20 units/§¡) 10 §¡
PCR Grade Water 1.25 §¢
º¸Á¸
-20µµ
°ü·Ã Á¦Ç°
- 5£§-exonuclease È°¼ºÀ» Á¦°ÅÇÑ PCR È¿¼Ò : TITANIUM® Taq DNA Polymerase (Code 639208)
- Æí¸®ÇÏ°í È¿°úÀûÀÎ hot start ¹ý : TaqStart Antibody (Code 639250)
- ¹ü¿ëÀûÀÎ agarose : SeaKem® LE Agarose (Code 50004)
- °íÈ¿À²ÀÇ Ligation premix ¿Í T-vectorÀÇ Á¶ÇÕ : Mighty TA-cloning Kit (Code 6028)
- Long strand DNAµµ methylated DNAµµ OK! °íÈ¿À² Competent Cell: E.coli HST08 Premium Competent Cells (Code 9128)
- ÄÄÆÑÆ®ÇÑ 96 well gradient PCR ±â±â : Clontech PCR Thermal Cycler GP

Keyword :

Mutagenesis
>
Error prone PCR¿¡ ÃÖÀû
Diversify PCR Random Mutagenesis Kit