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Ch9 DNA-Based Information Technologies - Coggle Diagram
Ch9 DNA-Based Information Technologies
Recombinant DNA
overexpression of proteins
transgenic food, animals...
molecular cloning of genes
DNA Cloning
用途
五大步驟
細節
Step1
Restriction Endonucleases
DNA Ligase:(1)通常用於DNA修復(2)人體中的DNA Ligase需要耗ATP才能作用(3)細菌中的DNA Ligase需要耗NAD才能作用
Step2
Antibiotic Selection
Cloning Vectors
.
BAC
YAC
PCR(Polymerase Chain Reaction)
Mix together(1)target DNA(2)primers(olidonucleotides complementary to target)(3)nucleotides:dATP,dCTP,dGTP,dTTP(4)thermostable DNA polymerase
Place the mixture into a thermocycler
Used to amplify DNA in the test tube
General steps of PCR
.
qPCR
Electrophoresis:for the seperation of DNA
Site-Directed Mutagenesis
DNA Fingerprinting
cDNA
Expression of cloned genes
Purification of Recombinant Genes
Fluorescence: it can be used to determine protein location In Vivo
GFP-Tagged Protein Localization
Immunofluorscence
DNA Microarrays
Photolitographic Synthesis of DNA-Containing Microships
Two-Color DNA Microarray Analysis on "Genome Chips"
Applications
Tissue-specific gene expression
Mutational analysis
Genome-wide genotyping
Single Nucleotide Polymorphisms(SNPs)
