DNA sequencing

yolajoy
Mind Map by , created over 5 years ago

BSc degree Bioinformatics Mind Map on DNA sequencing, created by yolajoy on 04/01/2014.

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yolajoy
Created by yolajoy over 5 years ago
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DNA sequencing
1 Maxan and Gilbert method
1.1 1 - double stranded DNA is labeled using radioactive phosphate (phosphate 32) at the 5' end of each strand
1.1.1 the attachment of Phosphate 32 is done using a polynucleotide kinase enzyme and 32P-dATP
1.2 2 - the double stranded DNA is treated with dimethyl sulphoxide and heated to 90 degree centigrade to separate the strands
1.2.1 these strands are then purified
1.3 3 - the strands are treated with various chemicals which cause base modification
1.3.1 formic acid modifies purines (A and G)
1.3.2 dimethyl sulphate modifies guanine
1.3.3 hydrazine modifies pyrimidines
1.3.3.1 adding NaCl prevents modification of thymines
1.4 4 - the altered/modified bases are removed, hot piperidine is used to cleave the sequence at modified bases
1.4.1 the remaining strands of DNA undergo gel electrophoresis, through these results we can work out what the DNA base sequence is

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2 Sanger method
3 next generation sequencing
3.1 sequencing by synthesis
3.2 sequencing by ligation
3.3 single molecule sequencing
4 is finding the order of nucleotides in a piece of DNA
4.1 sequencing can be done on anything which contains genetic information
4.1.1 animals
4.1.2 plants
4.1.3 bacteria
4.1.4 archaea
4.2 depending on the method used you may be provided with DNA or RNA sequences
5 sequencing can be used to determine the sequence of
5.1 individual genes
5.2 large genetic regions such as gene clusters or operons
5.3 chromosomes
5.4 entire genomes
6 the results of sequencing may be used by
6.1 researchers to further scientific progress
6.2 by medical proffessionals to aid treatment decisions and genetic counselling

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