genomic complexity
DESCRIPTION
Genomic Complexity. How many genes in the chromosome?. DNA or RNA? Double Stranded or Single Stranded? Shape? Number of copies? Size in base pairs? Assume 400 amino acids per protein, then how many bases per gene for a typical protein? How many genes possible? Do genes overlap? - PowerPoint PPT PresentationTRANSCRIPT
Genomic Complexity
How many genes in the chromosome?• DNA or RNA?• Double Stranded or Single Stranded?• Shape?• Number of copies?• Size in base pairs?• Assume 400 amino acids per protein, then how many bases
per gene for a typical protein?• How many genes possible?• Do genes overlap?• Are both strands transcribed?
Consider the Ribosome
And its operon
Studying Macromolecules –Extracting DNA
Cell Pellet
Random fragments ~20-40 kbp
Extraction of Chromosomal DNA
• DNeasy Tissue Kit• Chromosomal
DNA, ~ 20-40 kbp size fragments
• Kim Foster
Chromosomal DNA
Chromosomal DNA
RNA
RNA
23 kbp
Hybridization
Quantitative Hybridization
Liquid Scintillation Counters
Photomultiplier Tubes
log Scale
Electronic Noise
3H 14C 32P
Spike
Experiment 1 – Total 14C-Uracil
10 µg DNA
10 µg RNA
5 µg RNA washes off
5 µg RNA sticks
Experiment 2 – rRNA 14C-Uracil
10 µg DNA
10 µg rRNA
>99% rRNA washes off
35 ng rRNA sticks
Experiment 3 – tRNA 14C-Uracil
10 µg DNA
10 µg tRNA
>99% tRNA does not stick
5 ng tRNA sticks