Transcription of 제한효소 / Ligation을 이용한 Cloning - TaKaRa CMS
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제한효소 / Ligation을 이용한 Cloning - TaKaRa CMS
[ ]1. Insert DNA ( DNA )(a) DNA cutting DNA , plasmid vector Cloning . Hind lll BamH I double digestion . DNA ( 1 ) Hind lll 1 BamH I 1 10 x K buffer 2 dH2O up to 20 ( ) 37 C 1 . Agarose gel . (10 ) loading buffer . DNA gel . DNA , UV . gel DNA ( gel extraction kit ). insert DNA A (b) PCR Vector ligation insert primer 5 . primer insert DNA PCR . + 5 3 base nucleotide TaKaRa Ex Taq Hot Start Version PCR . 10 Ex Taq Buffer Mg2+ plus 5 dNTP Mixture mM 4 200 M primer 1 ~ M final conc.
18 · Life Science & Biotechnology 48 3. Insert DNA와 linear plasmid의 ligation DNA Ligation Kit〈Mighty Mix〉를 사용할 경우 ① 반응액을 조제한다.
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Introduction: Cloning (DNA Ligation) &, Introduction: Cloning (DNA Ligation) & Transformation, OF BACILLUS SUBTILIS sacB (LEVANSUCRASE), Ligation cloning, Cloning, LentiCRISPRv2 and lentiGuide-Puro: lentiviral, Biotechnology and Recombinant DNA, DNA sequencing methods – general principles of, And bacterial expression system optimization, KOD -Plus