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目录 The Principle and Application of Common Used Techniques in Molecular Biology chapter 18.

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Presentation on theme: "目录 The Principle and Application of Common Used Techniques in Molecular Biology chapter 18."— Presentation transcript:

1 目录 The Principle and Application of Common Used Techniques in Molecular Biology chapter 18

2 目录 1 Molecular Hybridization and Blotting Technique

3 目录 nucleic acid hybridization In DNA renaturation process, if the type of single-stranded DNA molecule in the same solution, the DNA or RNA together with, as long as between single-stranded DNA or RNA molecule with a base pairing relationship can between the different double-stranded hybrid molecules 1. Principles and blot hybridization techniques

4 目录 Refolding RNADNA

5 目录 Blot Various physical methods using gel electrophoretic transfer to NC biological macromolecules other film, making the solid phase molecule. This technique is similar to absorb the ink on the paper with a blotter, so called "blotting", translated blot technique.

6 目录 Radionuclide, biotin or a fluorescent dye labeled polynucleotide chain end or the whole chain of known sequence is called a "probe", the probe may bind to the immobilized polynucleotide of the NC membrane, determine whether homologous nucleic acid molecule. Probe Technology

7 目录 2. Category Blot and application (1) Southern blotting ( 2 ) Northern blotting ( 3 ) Western blotting Analysis of genomic DNA, recombinant plasmids and bacteriophages used. For qualitative and quantitative analysis of RNA. Quantitative and qualitative for protein interaction studies.

8 目录 Other : dot blotting in situ hybridization DNA array DNA chip

9 目录 Comparison of three kinds of Blot

10 Molecular hybridization experiments ① ② ③

11 目录 Autoradiograph

12 目录 DNA chip

13 目录 2 The Principle and Application of PCR Technology

14 目录 5 Primer 1 5 Primer 2 Cycle 2 Cycle 1 5 5 5 5 5 5 Template DNA 1 、 Principles of PCR technology 5 5 5 5 5 5 5 5

15 目录 Cycle 3 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 After 25 to 30 cycles, the content of the template DNA can be expanded 100 times or more.

16 目录 Schematic diagram of PCR technology

17 目录 The basic steps of PCR reaction 变性 95˚C 延伸 72 ˚ C 退火 Tm-5 ˚ C

18 目录 Template DNA Primers Thermostable DNA polymerase dNTPs Mg2 + The basic components of PCR system

19 目录 Real-time PCR technology principle Q R 3'3' 3'3' 5'5' 5'5' Upstre am primer Down strea m prime r Fluorescently labeled primers R Q 3' 3'3' 5'5' 5'5'

20 目录 Real-time PCR TaqMan probe Schematic

21 目录 3 Gene Library

22 目录 genomic DNA library cDNA library gene library Refers to an organism containing a DNA sequence of the entire population of clones.

23 目录 1. Genomic DNA Library Genomic DNA library of genomic DNA is the information on an organism (including all coding and non-coding region) in the form of storage of cloning a DNA fragment populations. Vector used to construct genomic libraries have phages, cosmids and yeast artificial chromosomes.

24 目录 Construction and screening of genomic libraries and cDNA library

25 目录 1 2 3 Examples of genomic library screening results

26 目录 cDNA library mRNA that contains a clonal population of all cells under certain conditions the expressed cDNA synthesized by reverse transcription of the sequence, it is stored in the form of a cDNA fragment of the gene expression of cells of information. 2. cDNA library

27 目录 4 Biological Chip Technique

28 目录 DNA 微阵列 (DNA microarray) 。 1. gene chip gene chip

29 目录 基因芯片工作流程示意图

30 目录 The protein molecules are highly dense arrangement of the lattice as a probe immobilized on a solid phase support material, when reacted with the target protein sample, the sample may be captured in the target protein, and then the target protein detection system for qualitative and quantitative analysis a technique. 2. Protein chips 蛋白质芯片 (protein chip)


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