sequencing gap - Rice Genome Annotation Project

More documents

Recommendations

Info

Construction of Transposon-MediatedLibraries• Transposons are genetic elementsthat provide mobile primer siteswithin the target DNA for<strong>sequencing</strong>• The GPS-1 <strong>Genome</strong> PrimingSystem (New England BiolabsE7100S) enables transposoninsertions by using TnsABC*transposase+• Only one random insertion occursper target DNATIGRTHE INSTITUTE FOR GENOMIC RESEARCH
Use of Transposon-mediated libraries:•Make a library using a transposon kit on a spanning clone.•Sequence the clones using the transposon primers•Assemble the sequencesPhysical map of thetransposon inserts:M13FM13FTransposon1 Transposon2N S N SRepeatClone DNAN1 S1M13RM13RN2S2TIGRFinal AssemblyTHE INSTITUTE FOR GENOMIC RESEARCH
Page 4 and 5:
Library Construction• Break the g
Page 6:
DNA Shearing with HydroShearHydroSh
Page 9 and 10:
Design Features of the BstXI Adapto
Page 11 and 12:
Template Production• Transform cl
Page 13 and 14:
Template Production LaboratoryCurre
Page 15 and 16:
Sequence Production LaboratoryCurre
Page 17 and 18:
Sample ElectropherogramTIGRTHE INST
Page 19 and 20:
What are phred quality values?The q
Page 21 and 22:
TIGRTHE INSTITUTE FOR GENOMIC RESEA
Page 23 and 24:
Assembling the fragmentsTIGRTHE INS
Page 25 and 26:
Merging two sequencesoverlap (19 ba
Page 27 and 28:
Forward-reverse constraints• The
Page 29 and 30:
Degenerate contigs• Too deep• T
Page 31 and 32: Scaffolding• Given a set of non-o
Page 33 and 34: Linking information• Overlaps•
Page 35 and 36: Assembly gapsphysical gapgroup Agro
Page 37 and 38: Problems with the data• Incorrect
Page 39 and 40: Unifying view of assemblyAssemblySc
Page 41 and 42: BAC-by-BAC Sequencing and Closure
Page 43 and 44: BAC Shotgun SequencingBAC DNASequen
Page 45 and 46: Why Completeness is Important• Im
Page 47 and 48: Levels of Genome ClosureTIGRTHE INS
Page 49 and 50: TIGRTHE INSTITUTE FOR GENOMIC RESEA
Page 51 and 52: Sequencing GapsContig AUnderlyingse
Page 53 and 54: Repetitive Areas• Repetitive area
Page 55 and 56: Mis-assembled repeatClones link dif
Page 57 and 58: Sequence editor• In this example
Page 59 and 60: Resultingcontig afterrepeatresoluti
Page 61 and 62: Collapsed Repeat-5.5 CE DipReadCove
Page 63 and 64: Sequence Validation: Editing• Fin
Page 65 and 66: Sequence Validation:Sequence covera
Page 67 and 68: Sequence Coverage in CloeTIGRTHE IN
Page 69 and 70: Clone Coverage ExampleCATCCGAAGACGA
Page 71 and 72: Finished BAC Ready for Annotation!B
Page 73 and 74: TIGRTHE INSTITUTE FOR GENOMIC RESEA
Page 75 and 76: HT TemplateLibraryConstructionHT Se
Page 77 and 78: Schematic View of 3 Typical Scaffol
Page 79 and 80: Short Tandem RepeatsTIGRTHE INSTITU
Page 81: Inverted RepeatsTIGRTHE INSTITUTE F
Page 85 and 86: Closure Challenges: SequencingThrou
Page 87 and 88: Homopolymeric tractsTIGRTHE INSTITU
Page 89 and 90: • Apply different sequencing chem
Page 91 and 92: Automated Finishing (cont’d)• L
Page 93 and 94: • Dead EndsBAC Project Challenges
Page 95 and 96: Filling small gaps in a chromosome
Page 97: Project Organization• Database to
show all

sequencing gap - Rice Genome Annotation Project

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?