ANTHROP 1AA3 Lecture Notes - Lecture 20: Eva Green, Elution, Capillary Electrophoresis
4 May 2016
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Lab 2: in lab two we have all our dna: lysis, binding to column, washing, elution. 10x buffer keep ph at level for polymerase mag. What is inhibition- to much dna/too little dna. Over come this but diluting our samples to remove inhibitors. Lab 4: check if pcr yieleded correct product. Different size move at different rates dna moves from cathode to positive anode. Can compare size to dna of known size (ladder) to figure out if correct product was formed. 5: why purify: getting rid of components from pcr and exchange buffers. Buffer exchange 10 x buffer to eb buffer same principle as extraction. 6: gel quantification must be at specific concentration for sequencing. Can compare intensity (how dark band is), and spatial comparision for size how far did the band run down the gel. Sanger sequencing by synthesis followed by capillary electrophoresis. Ddntps stop replication flueroscently labeled dntps- for synthesis chromatogram- output of sequencing bases are different colours.
