BC basic - from big bang - PowerPoint PPT Presentation

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BC basic - from big bang

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DNA1 DNA2 RNA Probe Southern hybridization Northern hybridization Juang RH (2004) BCbasics Preparation of Traditional Nucleic Acid Probe Amino acid sequence GLY-ASP ... – PowerPoint PPT presentation

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Title: BC basic - from big bang


1
Hybridization of Nucleic Acids
DNA1
DNA2
RNA
Probe
Southern hybridization
Northern hybridization
Juang RH (2004) BCbasics
2
Preparation of Traditional Nucleic Acid Probe

Codon degeneracy
The nucleic acid sequence is Deduced from amino
acid sequence
Synthesizing oligonucleotide
Chemical synthesis
PROBE GGGGACGAGTCCTCCGTTCT
Juang RH (2004) BCbasics
3
Probe is labeled with radioactive 32P
Hybridization
DNA denaturation
Target gene
Single colony
Lysed
Juang RH (2004) BCbasics
4
Colony Is Screened by Hybridization with Probe
Colony hybridization
Cover with filter paper
Transferring
Collect filter paper
Autoradiography
Dissolve cell DNA denatured
Add probe
Juang RH (2004) BCbasics
5
Biochip Based on Hybridization
Sample DNA
Complementary DNA hybridize
Biochip
Each spot contains known DNA
Signal appears
Schena (2000) Microarray Biochip Technology, p.
A31
Juang RH (2004) BCbasics
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