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<Biotechnology techniques>

<Biotechnology techniques>. 특정 유전자 발현 검토. PCR(polymerase chain reaction). ㅖㅊ. RT-PCR: 특정 mRNA. DNA Microarray(DNA chip): transcriptome. Applications of NGS. Whole genome (re)sequencing de novo genome assembly (much harder with shorter reads)

justin-kaufman
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<Biotechnology techniques>

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  1. <Biotechnology techniques>

  2. 특정 유전자 발현 검토

  3. PCR(polymerase chain reaction) ㅖㅊ

  4. RT-PCR: 특정 mRNA

  5. DNA Microarray(DNA chip): transcriptome

  6. Applications of NGS • Whole genome (re)sequencing • de novo genome assembly (much harder with shorter reads) • Variant detection (SNPs, indels) and copy number, structural variation • Trancriptome • Whole transcriptome sequencing • Gene expression profiling • Small RNA sequencing NGS Next Generation Sequencing • Targeted resequencing • Exon sequencing • Epigenome • Chromatin Immunoprecipitation sequencing (Chip-Seq) • Methylation analysis • -MeDIP(Methylated DNA Immunoprecipitation ) • :Anti m5C specific antibody 를 이용한 pull down • Unknown C methylation 영역 찾기에 유리

  7. Next Generation Sequencer: Three Leading Platforms With 3730s capillary, ~60 Mb per year

  8. Sanger sequencing: chain termination using dideoxynucleotides Source: http://www.bio.davidson.edu/Courses/Bio111/seq.html

  9. Pyrosequencing

  10. 2 D electrophoresis/MALDI-TOF MASS: proteome 분석 • 2-DE by subfractionation methods pH acidic  basic  MW Kidney tissue, homogenated post-mitochondrial protein, TCA/acetone, colloidal coomassie blue staining Kim-Chulhong, PBBRC

  11. Advantage of MALDI-TOF MS • 빠르다. 즉 이온화 및 분석물의 검출이 모두 밀리세컨드안에 끝난다. • 결과가 분명하다. 왜냐하면 근본적으로 질량 대 전하비(mass-to-charge ratio:m/z)에 근거하기 때문에 핵산의 2차 구조에 영향을 받는 전기영동이나 hybridization-array (e.g. DNA chip) 보다 정확하다. • 모든 과정, 시료의 준비에서 결과 처리까지의 과정이 완벽하게 자동화 가능하다. 따라서 MALDI-TOFMS는 시료의 대량분석 (high-throughput)의 가능성을 제공한다. Kim-Chulhong, PBBRC

  12. Flight Tube Detector 4-25 kV Ion Source Reflector (Ion Mirror) MALDI-TOF MS • Reflector Mode : isotope peak 분리가능 • Ion mirror (reflector)를 사용하여 비행거리를 증가시킴으로 resolution을 높인다 • Linear Mode : resolution 이 떨어진다. R L Liner mode Kim-Chulhong, PBBRC

  13. Trasfection

  14. Transgenic mice

  15. ㅏㅐ Knockout mice

  16. RNAi

  17. Footprinting (footprint analysis)

  18. Bandshift assay or Electrophoretic mobility shift assay (EMSA)

  19. Luciferase/reporter vector Promoter analysis

  20. Receptor binding assay

  21. Radioimmunoassay (RIA)

  22. Radioimmunoassay (RIA)

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