Lecture 13 Notes

Question 1

oligonucleotide primer (20 nucleotides)

Answer 1

the primer defines a fixed, common reference point for DNA strands synthesized in the sequencing reaction

Question 2

next gen sequencing

Answer 2

• various high throughput methods used to sequence random fragments of DNA without the need for making recombinant DNA clones
• genomic DNA randomly fragmented by shearing
• linker/adaptors ligated to ends

Question 3

human genome project

Answer 3

• human genome is 3 billion base pairs
• major accomplishments: DNA sequence of entire genome = RefSeq; large fragments of genome isolated and organized; DNA sequence annotated to define genes; DNA sequences of model organisms and other species

Question 4

Genome projects 1

Answer 4

determining sequence of human genome

• fragmenting the genome
• sequencing the genome
• reconstructing the genome sequence from fragments
• finding the genes
• what the genome looks like
• bioinformatics tools

Question 5

fragmenting the genome

Answer 5

• cut genomic DNA into fragments of different average sizes
• separate genomic DNA fragments into a particular size class
• insert the DNA fragments into cloning vectors to generate libraries

Question 6

paired end sequencing

Answer 6

preferred method for sequencing eukaryotic genomes for the first time