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3 Cards in this Set
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WGS-construction physical maps
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1)0.4-1.2 kb shotgun library
5-20 kb shotgun library~~~~~~~sequence production 2)genome wide raw sequences (6-8x) editing, contig assembling n gap filing |
from randomly shear n clone in M13 and/plasmids
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problem with repetitive sequences & how to overcome?
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interfere with genome assembly
can map to multiple location within genome so, assembled seq must remain broken at repetitive seq by paired end sequencing-bridge gap in assembly seq will be generated from both ends gDNA fragment of known size if paired end seq flank repetitive element,can allow assembly scaffold, contig sets physically linked by paired end sequencing |
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Clone by clone shotgun sequencing
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1.150kb BAC library - fingerprinting, BAC end sequencing
2.BAC based physical map select minimally overlapping BAC from map n make shotgun libraries from one BAC at time 3.produce 1kb shotgun library produce 5-10kb shotgun library then,sequence production one BAC at a time 4. individual BAC raw seq (6-8x) editing, contig assembling, gap filing, n entire BAC sequence extension |
from partially digest n clone in BAC
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