鸟枪法测序原理

Click to edit Master title style,Click to edit Master text styles,Second level,Third level,Fourth level,Fifth level,*,Regional mapping,Regional mapping,Minimal tiling path selected for sequencing.,Regional mapping,10/15/2024,20 kbp,300 bp,Molecular weight,marker every,5th lane,Restriction,fragment,fingerprinting,- BAC clones are grown,in 96-well format,- Hind III digest,- 1% agarose,Contig assembly,Clone,A,B,C,D,E,F,G,FPC*,Overlap identification by,restriction pattern similarities,Facilitated contig assembly,*,Sanger,Centre,C. Soderlund, I Longden and R. Mott,*,*,*,*,*,*,All restriction fragments within,a clone selected for the tiling,path must be verified by their,presence in overlapping clones.,: vector fragments,: insert fragments,BCM-,HGSC,Shotgun Sequencing I :RANDOM PHASE,Bac,Clone:,100-200 kb,Sheared DNA:,1.0-2.0 kb,Sequencing,Templates:,Random,Reads,BCM-,HGSC,Shotgun Sequencing II:ASSEMBLY,Consensus,Sequence,Gap,Low Base,Quality,Single,Stranded,Region,Mis,-Assembly,(,Inverted,),BCM-,HGSC,Consensus,Sequence,Gap,Low Base,Quality,Single,Stranded,Region,Mis,-Assembly,(,Inverted,),BCM-,HGSC,Shotgun Sequencing III: FINISHING,Consensus,Sequence,Gap,Single,Stranded,Region,Mis,-Assembly,(,Inverted,),BCM-,HGSC,Shotgun Sequencing III: FINISHING,Consensus,Sequence,Gap,Mis,-Assembly,(,Inverted,),BCM-,HGSC,Shotgun Sequencing III: FINISHING,Consensus,Mis,-Assembly,(,Inverted,),BCM-,HGSC,Shotgun Sequencing III: FINISHING,BCM-,HGSC,Shotgun Sequencing III: FINISHING,High Accuracy Sequence:,100,000?,- Base Quality High or Low?,BCM-,HGSC,-,Mis,-Assemblies?,- Duplications?,- Regions not covered,- Regions very densely covered,Complete an accurate, high quality sequence,of the human,genome,by the end of 2003,.a,working draft,can be completedwithin,the next three years,BCM-,HGSC,That (draft) sequence will be of lower accuracy and contiguity.,will be useful for finding genesand other features.,BCM-,HGSC,Working Draft Sequence Options:,0.5 -1.0 X (2 reads/kb) - Skimming,3.5 - 4.0 X (9 reads/kb) -half-shotgun,6.5 - 8.0 X (18 reads/kb) - pre-finished,10 X (22-24 reads/kb) - deep shotgun,