Transcript Hybridization of Nucleic Acids RNA DNA1 DNA2

Hybridization of Nucleic Acids
DNA1
DNA2
RNA
Probe
Northern
hybridization
Southern
hybridization
Juang RH (2004) BCbasics
Preparation of Traditional Nucleic Acid Probe
Amino acid sequence
GLY-ASP-GLU-SER-SER-VAL-LEU----GGG-GAC-GAG-TCC-TCC-GTT-CTC---
*
*
*
*
Nucleic acid sequence
The nucleic acid sequence is
Deduced from amino acid sequence
Chemical synthesis
*
*
*
* Codon degeneracy
Synthesizing
oligonucleotide
PROBE: GGGGACGAGTCCTCCGTTCT
Juang RH (2004) BCbasics
Probe is labeled with radioactive 32P
32
P
GG
G GAC G
AG
Hybridization
TC
GG
T
C
TC
CG
A
T
T
G
T
C
T
C
C
C
C
T
CAG
G
G
TCC
A
A
G
G
AGGCAA
C
C
T
A
DNA
denaturation
Target gene
Single colony
Lysed
Juang RH (2004) BCbasics
Colony Is Screened by Hybridization with Probe
Colony hybridization
Transferring …
Collect filter
paper
Dissolve cell
Autoradiography
DNA denatured
Add probe
Juang RH (2004) BCbasics
Cover with
filter paper
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