Lecture 8 LC710- 1st + 2nd hr

Lecture 8 LC710- 1st + 2nd hr - Hybridization of Nucleic...

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Hybridization of Nucleic Acids RNA DNA1 DNA2 Probe Southern hybridization Northern hybridization Juang RH (2004) BCbasics
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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 * Codon degeneracy * * * * * * * Synthesizing oligonucleotide PROBE: GGGGACGAGTCCTCCGTTCT PROBE: GGGGACGAGTCCTCCGTTCT The nucleic acid sequence is Deduced from amino acid sequence Chemical synthesis Juang RH (2004) BCbasics
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G G A T C C A T C C C C TG C T C A G G A G GCA A G A G G C A T P 32 Target gene DNA denaturation Single colony Lysed Hybridization Probe is labeled with radioactive 32 P Juang RH (2004) BCbasics
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Colony Is Screened by Hybridization with Probe Cover with filter paper Autoradiography Add probe Transferring … Collect filter paper Dissolve cell DNA denatured Juang RH (2004) BCbasics Colony hybridization
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Sample DNA Each spot contains known DNA Biochip Schena (2000) Microarray Biochip Technology, p. A31 Complementary DNA hybridize Signal appears Biochip Based on Hybridization Juang RH (2004) BCbasics
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The Genetic Code Initiation and termination Codons Initiation codon: AUG Termination codons: UAA, UAG, UGA Degeneracy: partial and complete Ordered Nearly Universal (exceptions: mitochondria and some protozoa)
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Key Points Each of the 20 amino acids in proteins is specified by one or more nucleotide triplets in mRNA. (20 amino acids refers to what is attached to the tRNAs!) Of the 64 possible triplets, given the four bases in mRNA, 61 specify amino acids and 3 signal chain termination. (have no tRNAs!)
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Key Points The code is nonoverlapping, with each nucleotide part of a single codon, degenerate, with most amino
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This note was uploaded on 02/14/2012 for the course BIO 710L taught by Professor Goldfarb during the Fall '10 term at Iowa State.

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Lecture 8 LC710- 1st + 2nd hr - Hybridization of Nucleic...

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