DNA extraction 분자생물학실험 SUBJECT

Sequence blast Restriction enzyme Mini-prep E.coli transformation TA Ligation PCR DNA EXTRACTION

Arabidopsis thaliana 식물체가 작아서 plant biology 분야에서 model organism 으로 널리쓰임 member of the mustard (Brassicaceae) family 유전학과 분자생물학의 기본 연구들에 주요한 장점을 제공한다 125Mb 중 115Mb 의 Approximately 115 Mb of the 125 Mb genome has been sequenced and annotated (Nature, 408: ; 2000). Extensive genetic and physical maps of all 5 chromosomes are available. The life cycle is short--about 6 weeks from germination to seed maturation. Seed production is prolific and the plant is easily cultivated in restricted space. Transformation is efficient utilizing Agrobacterium tumefaciens. A large number of mutant lines and genomic resources is available. Such advantages have made Arabidopsis a model organism for studies of the cellular and molecular biology of flowering plants. 3 분자생물학실험 Materials & Method

분자생물학실험 Samples(Arabidopsis) Extraction buffer, Isopropanol, 70% Ethanol, D.W. pipette, tip, E-tube, rack, grinder, centrifuge, PCR DNA extraction buffer 200mM Tris-Cl (pH7.5) 250mM NaCl 25mM EDTA 0.5% SDS(Sodium dodecyl sulfate)

Materials & Method 분자생물학실험 DNA isolation 1. Grind samples and add 500ul DNA extraction buffer. 2. Centrifuge at 13,000 rpm for 5min. 3. Mix Supernatant 400ul + isopropanol 400ul (1:1) 4. Centrifuge at 13,000 rpm for 20min. 5. Discard supernatant and add 70% Ethanol 500ul 6. Centrifuge at 13,000 rpm for 5min. 7. Dry the DNA pellet 8. Dissolve DNA in D.W.

Materials & Method 분자생물학실험 Isopropanol

DNA extraction buffer 에 들어가는 시약의 역할, 왜 필요한지 ? 원리 DNA extraction buffer ( 아래 표 완성 ) Result & Discussion 분자생물학실험 ReagentFinal conc.Stock conc.Volume Tris-Cl(pH 7.5)200mM1M ㎖ NaCl250mM5M ㎖ EDTA25mM0.5M ㎖ SDS0.5%10% ㎖ D.W-- ㎖ Total 250 ㎖ 표 아래에 계산과정 논리적으로 정리할 것 Ex) 1M NaCl=58.44gNaCl

다음 실험 레포트 분자생물학실험 -DNA extraction -> Material & Method ->Results& Discussion Report -PCR Primer design, PCR 예비 레포트 -9 월 16 일 오후 6 시까지 제출하기