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702281
DNA sequencing
Description
BSc degree Bioinformatics Mind Map on DNA sequencing, created by yolajoy on 01/04/2014.
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bioinformatics
bioinformatics
bsc degree
Mind Map by
yolajoy
, updated more than 1 year ago
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Created by
yolajoy
over 10 years ago
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Resource summary
DNA sequencing
Maxan and Gilbert method
1 - double stranded DNA is labeled using radioactive phosphate (phosphate 32) at the 5' end of each strand
the attachment of Phosphate 32 is done using a polynucleotide kinase enzyme and 32P-dATP
2 - the double stranded DNA is treated with dimethyl sulphoxide and heated to 90 degree centigrade to separate the strands
these strands are then purified
3 - the strands are treated with various chemicals which cause base modification
formic acid modifies purines (A and G)
dimethyl sulphate modifies guanine
hydrazine modifies pyrimidines
adding NaCl prevents modification of thymines
4 - the altered/modified bases are removed, hot piperidine is used to cleave the sequence at modified bases
the remaining strands of DNA undergo gel electrophoresis, through these results we can work out what the DNA base sequence is
Attachments:
Maxam and Gilbert
Sanger method
next generation sequencing
sequencing by synthesis
sequencing by ligation
single molecule sequencing
is finding the order of nucleotides in a piece of DNA
sequencing can be done on anything which contains genetic information
animals
plants
bacteria
archaea
depending on the method used you may be provided with DNA or RNA sequences
sequencing can be used to determine the sequence of
individual genes
large genetic regions such as gene clusters or operons
chromosomes
entire genomes
the results of sequencing may be used by
researchers to further scientific progress
by medical proffessionals to aid treatment decisions and genetic counselling
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