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Sheet1 - ALEXA

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Performs Transcript Expression Gene

Tool split-read reconstruction Analysis expression

alignment (assembly) (any) analysis





Alexa-Seq N N Y Y





Cufflinks N Y Y Y





Scripture N Y Y N





SpliceMap N N Y Y





TopHat Y N N N





MMES Y N N N





G-Mo.R-Se Y Y N N





SplitSeek Y N N N





GSNAP Y Y N N





* Transcript expression values are non-parsimonious

** Not supported (current version not stable)







Performs Transcript Expression Gene

Tool split-read reconstruction Analysis expression

alignment (assembly) (any) analysis





Alexa-seq N N Y Y





Cufflinks N Y Y Y

Scripture N Y Y N





SpliceMap N N Y Y





TopHat Y N N N





MMES Y N N N





G-Mo.R-Se Y Y N N





SplitSeek Y N N N





GSNAP Y Y N N





* Transcript expression values are non-parsimonious

** Not supported (current version not stable)

Quantitative

Transcript specific Exon junction Expression level

alternative expression

expression analysis expression sensitivity

analysis





Y Y Y Junction





Y N Y Transcript





N* N N Transcript*





N Y N Junction





N Y N Junction





N Y N Junction





N N N Transcript*





N Y N Junction





N N N N/A









Quantitative

Transcript specific Exon or Junction Expression level

alternative expression

expression analysis expression sensitivity

analysis





Y Y Y Junction





Y N Y Transcript

N* N N Transcript*





N Y N Junction





N Y N Junction





N Y N Junction





N N N Transcript*





N Y N Junction





N N N N/A

Minimum read length

Output required or

recommended



Expression and structure information for junctions and

No minimum (tested on

genes; UCSC track info, extensive alternative expression

36bp-100bp reads)

visualization / statistics / graphs



Transcript information and expression statistics, BED, GTF 75bp





Transcript structure information and non-parsimonious

75bp

expression statistics, BED





Alignments. SAM, BED, Wig 50bp





Alignments. SAM, BED, Wig 75bp





Identified splice junctions and p-values 25bp





Transcript structure information. GFF 25bp





Alignments. BED 50bp





Minimum 14bp (tested on

Alignments. SAM and FASTA

36bp reads)









Minimum read length

Output required or

recommended



Expression and structure information for junctions and

No minimum (tested on

genes; UCSC track info, extensive alternative expression

36bp-100bp reads)

visualization / statistics / graphs



Transcript information and expression statistics, BED, GTF 75bp

Transcript structure information and non-parsimonious

75bp

expression statistics, BED





Alignments. SAM, BED, Wig 50bp





Alignments. SAM, BED, Wig 75bp





Identified splice junctions and p-values 25bp





Transcript structure information. GFF 25bp





Alignments. BED 50bp





Minimum 14bp (tested on

Alignments. SAM and FASTA

36bp reads)

Performs comparisons Relevant

Visualization tool between conditions (ex. comparison to

tumor vs normal) Alexa-seq



Extensive, includes

custom graphs and links Y -

to UCSC browser



UCSC browser Y Y





UCSC browser N N





UCSC browser N N





N N N





N N N





Grape Genome Browser N N





UCSC browser N N





UCSC browser N N









Ability to identify

rearrangements / Junction Identification Implementation

indels





N Database Perl/R/Unix





N Predicted from data C++

N Predicted from data Java





N Predicted from data Python





N Predicted from data C++/Python





Published algorithm

N Predicted from data

only





N Predicted from data Perl





Y Predicted from data Perl



Source code in C,

Database or predicted form

Y utility programs in

data

Perl

Data type

Citation

supported







Illumina In preparation



Trapnell et al., 2010,

Paired-end

Nat. Biotech., 28:511-

Illumina reads

515

Guttman et al., 2010,

Illumina Nature Biotech, 28:503-

510 |

http://www.nature.com/

Au et al., 2010, NAR,

Illumina DOI

10.1093/nar/gkq211 |

http://www.stanford.ed

Trapnell C, Pachter L,

Illumina Salzberg SL. 2009,

Bioinformatics,

Wang et al., Plos |

25(9):1105-1111One,

Illumina 2010, Vol 5, Iss 1,

e8529

Denoeud et al., 2008,

Illumina Genome Biol,

9(12):R175 |

http://genomebiology.co

Ameur et al.2010,

SOLiD only Genome Biol.,

11(3):R34. |

Illumina | sodium http://genomebiology.co

Wu and Nacu, 2010,

bisulfite-treated Bioinformatics,

DNA sequencing 26(7):873-881 |

(for analysis of http://bioinformatics.oxf









Validated

Public tool Open Source

events



88% of 192

Y Y (GPL) alternativel

y expressed

exon by

5/5

Y Y alternativel

y spliced

genes by RT-

90% of

Y Unknown downstream

alternative

5′ startof all

87.9%

Y Unknown (151,317)

junctions,

and 41.2%

Y Unknown None



20/20 false

N N/A positives

(FP)

validates,

175

Y** Y (18.5%) of

944 cDNA

non-intron-

2 exon

Y Y (GPL) junctions in

2 genes



Y Unknown None

Validation method







RT-PCR, qPCR, Sanger Sequencing, mRNA/EST alignments, Splicing microarrays





RT-PCR | EST and RefSeq alignments | ChIP-seq

H3K4me3 marks on alternative promoters | polyadenylation motifs supporting alternate 3' ends |

RT-PCR of novel exons followed by Sanger sequencing





EST mapping | RT-PCR





N/A





RT-PCR for FP, EST support for FN





comparison to cDNA library from same tissues: 944 events detected in cDNA librarys vs 1,602

events detected by G-Mo-R-Se = 175 events in common.





EST and gene prediction support





N/A

Performs comparisons

Visualization tool between conditions (ex.

tumor vs normal)



Extensive, includes custom graphs and links to UCSC

Y

browser





UCSC browser Y

UCSC browser N





UCSC browser N





N N





N N





Grape Genome Browser N





UCSC browser N





UCSC browser N

Practical

Relevant comparison to Alexa-seq comparison to

Alexa-seq





- -





Y Y

N N





N N





N N





N N





N N





N N





N N

Organism and

Tissue Samples

Data type supported Citation

(for publicly

available analyses

to date)

Human (37

libraries) / Mouse (4 Illumina In preparation

libraries)

Mouse myoblast cell Trapnell et al.,

line at multiple Paired-end Illumina reads 2010, Nat.

differentiation Biotech.,

timepoints 28:511-515

Guttman et al.,

Mouse ESCs, NPCs,

Illumina 2010, Nature

ATCCs

Biotech,

28:503-510 |

Au et al., 2010,

Human brain

Illumina NAR, DOI

samples (4)

10.1093/nar/gk

q211 | C,

Trapnell

Mouse brain Illumina Pachter L,

Salzberg SL.

Mouse brain, liver, 2009, et al.,

Wang

muscle; Human Illumina Plos One, 2010,

embryonic kidney Vol 5, Iss 1,

and B-cell lines e8529

Denoeud et al.,

Grape leaf, root,

Illumina 2008, Genome

stem, callus

Biol,

9(12):R175 |

Ameur et

Mouse oocytes (2) SOLiD only al.2010,

Genome Biol.,

Human UHR from Illumina | sodium bisulfite-treated DNA 11(3):R34. |

Wu and Nacu,

MAQC, simulated sequencing (for analysis of methylation 2010,

reads from genome status) Bioinformatics,

with introduced 26(7):873-881

URL







www.alexaplatform.org





http://cufflinks.cbcb.umd.edu/

http://www.broadinstitute.org/software/scripture

/



http://www.stanford.edu/group/wonglab/SpliceM

ap/





http://tophat.cbcb.umd.edu/





N/A





http://www.genoscope.cns.fr/externe/gmorse/





http://solidsoftwaretools.com/gf/project/splitseek





http://research-pub.gene.com/gmap/



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