140 likes | 387 Views
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?
E N D
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?
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 23 kbp Chromosomal DNA RNA Chromosomal DNA RNA
Liquid Scintillation Counters Photomultiplier Tubes Electronic Noise Spike 3H 14C 32P log Scale
Experiment 1 – Total 14C-Uracil 10 µg RNA 5 µg RNA sticks 10 µg DNA 5 µg RNA washes off
Experiment 2 – rRNA14C-Uracil 10 µg rRNA 35 ngrRNA sticks 10 µg DNA >99% rRNA washes off
Experiment 3 – tRNA14C-Uracil 10 µg tRNA 5 ngtRNA sticks 10 µg DNA >99% tRNA does not stick