PCL102H1 Lecture Notes - Lecture 7: Sanger Sequencing, Microbead, Human Genome Project
12 Feb 2016
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Week #3: lecture 7: sequencing genomes with next generation sequencing. Challenges of sequencing: throughput, scalability, resolution and speed. Use dna polymerase to make new dna. Result are made by process of electrophoresis. Emit fluorescent light specific to base pair (allows detection) Separation by charge dna fragment of different lengths; last base pair indicated by type of florescent dye. Computer reads the results and produces an image; time and labour saving. Dna is only sequencing 1-2 thousand bases at a time. Order of fragments needed to understand genome. Top down approach (traditional) sequences are kept in order as they are cut (time consuming) Faster, cheaper, fragments sequenced with no attention to order. Computer uses the overlaps in sequences to determine order. Human genome project was done this way* Dna cut into fragments and separated into single strands. Microbeads immersed in emulsion oil with reagents to amplify dna. Each bead is placed in a micro well on a microplate.
