BIOL 200 Lecture Notes - Lecture 14: Apyrase, Luciferase, Phi X 174
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1977: first sequenced genome (bacteriophage phix174) mtdna: polycistronic. Assembly: reconstruct the original sequence using overlapping fragments (find the overlaps, align) Form scaffold: consists of contigs, w/ gaps of known length. Dna isolation --> fragmentation --> library --> amplification --> sequencing --> assembly. Illumina sequencing: all 4 dntps added in an attempt to pair the template base, simultaneously. This way it"s more efficient, and the sequencing would make fewer mistakes when there"s competition b/w the dntps. (think about the video!) Similar to sanger: nucleotides that act as terminators. Use reversible terminators (can be removed and reused. You just keep track of what colors were produced. ) Pacific biosystems: (pacbio) each nucleotide is fluorescent; fluorescence not coupled to base; coupled to the. Think about the video with the dna pol. Use a dna pol that, as it works, shows diff. colors of fluorescence from those fluorescent phosphate- linked nucleotides.
