BIOL114 - Lecture 4 - Genomics

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  • Created by: Katherine
  • Created on: 15-03-16 16:47
What are the aims of the Human Genome Project?
To determine the sequences of the 3 billion chemical base pairs that make up human DNA, identify all the genes in human DNA, improve tools for data analysis, store this information in databases making it available to researchers, address issues
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What type of issues could be addressed?
Ethical, legal and social issues that may arise from the project.
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When was the draft of the human genome produced?
In 2001, but we still don't have a complete sequence, so it is how far we've come since then. Quite often you can get gaps in the sequence. There are some parts which are more difficult to sequence than others.
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How many approaches are there to genome sequencing?
2
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What is Hierarchical genome sequencing?
Map based - based on clones. It was done on BAC-b-BAC vector. you construct a linkage map,
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What does BAC stand for?
Bacterial artificial chromosome. It is a vector that take bigger bits of DNA, and so is useful when cloning genomes.
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What are the steps of hiereachical genome sequencing?
Construction of genetic linkage map, construction of ordered clone maps, sequencing of ordered clones.
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What are genes on a cytogenetic map located by?
FISH - Fluorescnet insitu hybridisation - the ability to use a flurosent tag to identify a gene while it is still on the chromosome.
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What do you do in Hierarchical sequencing?
Locate the genes on the chromosome, you do mapping by looking at recobination frequencies and then order and sequence the clones.
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What is Whole genome shotgn sequencing?
Taking the whole of a genome and cutting it into short segments which are cloned in plasmids and sequenced using computer software.
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What is next generation sequencing?
Using a single template strand and amplifying this to produce an enormous number of identical ifragments. They are sequenced in parallel (at the same time)
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What are the stages of next generation sequencing?
Genomic DNA is fragmented, each fragment is isolated with a bead, using PCR, copies of a fragment are made, each attach to the bead by 5' end. The bead is placed into a well with DNA polymerases and primers. Nucleotides added. Repeated.
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What can you do with a genome sequence?
Store the sequence in a database e.g. NCBI Genbank, Identify all genes, compare with other species (comparatice and evolutionary genomic), post-genomic (measure expression (microarrays) and test function of all genes.
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Why is bioinformatics important?
It finds computational methods for storing, manipulating and analysing huge amounts of data.
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How to identify genes?
You could look for an open reading frame (this is where the gene starts and ends). If you look for these particular tags, you can see if you can find the beginning and end of a gene. Then you can compare this to other known genes. You use BLAST .
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How many types of BLAST programmes are there?
2 - BLASTN (nucleotide quer vs nucleotide database), BLASTP - proteiin query vs protein database.
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There are conventions about how BLASTP outputs are written, what is this?
In units of 60 normally
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What is repetitive DNA?
Chunks of DNA that just keep appearing in the genome, some of it is quire simple (SSR) some of it are very large duplications (we don't know if this is the combination of organisms, or a fluke that has replicated over time).
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Is complexity in gene numbers?
No
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How does complecity come about?
It's about the combination, not just gene number.
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What can you do with the mapping of the chromosomes?
Comparisons
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What are the practical benefits of the genome project?
Molecular medicine (identification of disease genes), energy sources and environemtnal applications, bioarachaeology, evolution, DNA forensics, Agriculture and breeding.
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What type of issues could be addressed?

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Ethical, legal and social issues that may arise from the project.

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When was the draft of the human genome produced?

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How many approaches are there to genome sequencing?

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What is Hierarchical genome sequencing?

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