Biology 1002B Lecture Notes - Lecture 22: Open Reading Frame, Dna Microarray, Sanger Sequencing
Document Summary
Lecture 20 current trend in cost of dna sequencing. Decreasing fast steps in typical dna sequencing methodologies. Get dna, purify it, get rid of proteins, rna. Break the dna up because cannot be sequenced from one end to another, has to be done in fragments. After sequencing it is assembled back: purification, fragmentation, amplification, sequencing fragments, assembly of fragment sequences structural features of dideoxynucleotides. They do not have a 3"oh, instead have h on the 3" carbon so no further base can be added basic mechanism of classical vs. automated sanger sequencing. Classical sanger sequencing involves 4 tubes, each tube has all four deoxynucleotides (datp, dttp, dgtp, dctp) and one of 4 dideoxynucleotides. Dna polymerase synthesizes the new strand until a dd nucleotide is incorporated which acts as a terminator. 4 tubes are run in 4 different lanes on the gel to distinguish between what dd nucleotide terminates the sequence.
