Department of Biotechnology Life Science Experiments 생 명 과 학 실 험생 명 과 학 실 험 특성화학부 생명공학과 식물분자생물학 실험실 6. PCR, Enzyme digestion SUBJECT 6. PCR, Enzyme digestion.

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Department of Biotechnology Life Science Experiments 생 명 과 학 실 험생 명 과 학 실 험 특성화학부 생명공학과 식물분자생물학 실험실 6. PCR, Enzyme digestion SUBJECT 6. PCR, Enzyme digestion

Department of Biotechnology Life Science Experiments 생 명 과 학 실 험생 명 과 학 실 험 특성화학부 생명공학과 식물분자생물학 실험실 6. PCR, Enzyme digestion Objective -Plasmid DNA isolation -Lysozyme boiling mini-prep -DNA quantification

Department of Biotechnology Life Science Experiments 생 명 과 학 실 험생 명 과 학 실 험 특성화학부 생명공학과 식물분자생물학 실험실 4.Mendelian segregation DNA Quantification ① 측정하고자 하는 DNA 용액을 일부를 채취하여 원하는 농도로 희석한다. ② 희석한 DNA 용액을 UV Spectrophotometer 에서 260nm 에서 absorbance 를 측정한다. ③ 측정한 absorbance 를 이용 DNA 농도와 양을 계산한다. UV Absorbance x DNA concentration factor x dilution factor = ? ng/ul Ex) x 50 x 100 = 4825ng/ul = 4.825ug/ul 4.825ug/ul x 40ul = 193ug

Department of Biotechnology Life Science Experiments 생 명 과 학 실 험생 명 과 학 실 험 특성화학부 생명공학과 식물분자생물학 실험실 6. PCR, Enzyme digestion Result Sample # Concentration(ng/ul)Total Quantity 1

Department of Biotechnology Life Science Experiments 생 명 과 학 실 험생 명 과 학 실 험 특성화학부 생명공학과 식물분자생물학 실험실 pDNA (15ng) (F +R)primer 2.5ul dNTP 2ul (10x) buffer 2ul Taq polymerase 1ul D.W Total 20ul (PCR) 6. PCR, Enzyme digestion

Department of Biotechnology Life Science Experiments 생 명 과 학 실 험생 명 과 학 실 험 6. PCR, Enzyme digestion 특성화학부 생명공학과 식물분자생물학 실험실

Department of Biotechnology Life Science Experiments 생 명 과 학 실 험생 명 과 학 실 험 특성화학부 생명공학과 식물분자생물학 실험실 (Enzyme digestion) DNA (300ng) 10XM 1 HindIII 1 PstI 1 D.W 10ul 6. PCR, Enzyme digestion

Department of Biotechnology Life Science Experiments 생 명 과 학 실 험생 명 과 학 실 험 HvTIP3;1prm(HindIII,PstI) 2,116 bp HindIII (2) PstI (2330) NdeI (2001) SphI (626)SphI (2046) KpnI (561)KpnI (1632) BamHI (290) BamHI (1363) BamHI (1613) SacII (219)SacII (692) SacII (1969) SacII (2142) pMD20-HvTIP3;1 Promoter (4,852bp)

Department of Biotechnology Life Science Experiments 생 명 과 학 실 험생 명 과 학 실 험 특성화학부 생명공학과 식물분자생물학 실험실 다음 실험 예비 레포트 7. Electrophoresis