Human genome project

• What were the aims of the human genome project

  To sequence the entire human genome and some non-human organisms.
• What were the two sequencing techniques used in the human genome project

  Hierarchical shotgun and whole-genome shotgun
• Who used whole-genome shotgun sequencing

  Celera genomics
• Which technique did the international human genome project use

  Hierarchical shotgun sequencing
• Why are plasmids not used in hierarchial sequencing

  The DNA fragments range from 150 - 350 kb which is too large.
• Which technique for the human genome project allowed repeat sequences to be sequenced

  Hierarchial shotgun
• What is a BAC

  This is a bacterial artificial chromosome which is man-made.

  It allows the incorporation of large DNA fragments which can replicate within a bacterial cell.
• How many tomes do BACs replicate

  once
• What is a collection of BACs called

  A BAC library
• What are the steps in hierarchical genome sequencing

  The DNA is fragmented.

  DNA is then cloned into BACs

  Ordered clone maps are created.

  Ordered clones are sequenced.
• Describe the steps in whole-genome sequencing

  DNA is fragmented.

  Fragments are cloned into vectors such as plasmids.

  Each fragment is then sequenced.

  Sequences are ordered.
• Which human genome sequencing technique was privately funded

  Whole genome shotgun
• Which human genome sequencing technique was publicly funded

  Hierarchial shotgun
• Which human genome sequencing technique ordered the fragments

  Hierarchial shotgun
• What are the two types of next generation sequencing

  Illumina and Ion torrent
• Briefly describe illumina sequencing

  DNA libraries are made using PCR. Each fragment is attached to a solid support prior to sequencing.
• Describe Ion torrent sequencing

  The DNA is fragmented and attached to a bead.

  PCR amplifies the fragments. The fragments attach via the 5 end to the bead.

  The beads are added to a solution of deoxyribonucleotide triphosphate and then washed.

  The process is repeated and the PCR will incorporate one of the deoxyribonucleotide triphosphates.
• Describe the positive result of ion torrent sequencing

  If one of the dNTPs is incorporated into the DNA, this changes the voltage, releasing phosphorous. And a flash of light is recorded.

  the order in which the dNTPs are added is recorded so you can determine the identity of which one was incorporated.
• What is the largest portion of DNA

  Repetitive DNA
• What is the smallest portion of DNA

  Exons