Difference between revisions of "Xenopus Genome Project"

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The [http://www.bio.utexas.edu/faculty/wallingford/ Wallingford] and Marcotte labs have obtained funding from the [http://www.ti3d.utexas.edu/ Texas Institute for Drug and Diagnostic Development] (TI3D) to begin sequencing of the ''X. laevis'' genome. We are primarily working with [https://wikis.utexas.edu/display/GSAF/About+Us Scott Hunicke-Smith] at the [https://wikis.utexas.edu/display/GSAF/Home+Page University of Texas Genome Sequencing and Analysis facility], with funding sufficient for ~20x coverage of the ''X. laevis genome'' using ABI SOLiD next-generation sequencing.  
 
The [http://www.bio.utexas.edu/faculty/wallingford/ Wallingford] and Marcotte labs have obtained funding from the [http://www.ti3d.utexas.edu/ Texas Institute for Drug and Diagnostic Development] (TI3D) to begin sequencing of the ''X. laevis'' genome. We are primarily working with [https://wikis.utexas.edu/display/GSAF/About+Us Scott Hunicke-Smith] at the [https://wikis.utexas.edu/display/GSAF/Home+Page University of Texas Genome Sequencing and Analysis facility], with funding sufficient for ~20x coverage of the ''X. laevis genome'' using ABI SOLiD next-generation sequencing.  
 
= CHORI-219 BAC sequencing =
 
We have started the first runs by sequencing 96 BACs from the [http://bacpac.chori.org/library.php?id=323 CHORI-219] library (vector: [http://www.sanger.ac.uk/Teams/Team53/psub/sequences/pbacgk.shtml pBACGK1.1]) at ~100X coverage. The selected BACs include ~70 genes of interest (Shroom3, Wnt5a, Glypican-4, Noggin, Gremlin, Pax6, Formin, etc., as initially identified by the group of [http://www.jgi.doe.gov/whoweare/cheng.html Jan-Fang Cheng] via probing the CHORI-219 library), as well as 10 BACs that have already been sequenced by the [http://www.jgi.doe.gov/ DOE Joint Genome Institute]/[http://www.hudsonalpha.org/genome-sequencing-center HudsonAlpha Genome Sequencing Center] to serve as positive controls for the sequencing and assembly pipeline. 
 
* CHORI-219 BACs: [[xdata:BACsFor1Percent.xls| List of 96 test BACs]] (MS Excel file)
 
 
See [[/XENLA_SA09023]] for more details. Three mate paired libraries were sequenced:
 
* X_laevis_WG - the ''X. laevis'' whole genome library, 5kb insert size - about 4.4GB raw data, 0.4GB high quality data<br>
 
* X_laevis_2kb - The set of 96 BACs, with 2kb insert size - about 3.6GB raw data, 0.3GB high quality data<br>
 
* X_laevis_5kb - The set of 96 BACs, with 5kb insert size - about 2.8GB raw data, 0.2GB high quality data<br>
 
This (very roughly) corresponds to >600X coverage by raw data, ~50X coverage by high quality data, of the BAC set.
 
* Given that we currently see better mapping of the shotgun SA09023 reads to ''X. tropicalis'' than to ''X. laevis'' (both to BACs and mRNAs), we're confirming the sample identity before continuing with whole genome sequencing. See the 'sanity check' [[/Species_Identification]] for details.
 
 
= J-strain whole genome sequencing =
 
In addition, we are generating several mate pair libraries of different sizes from genomic DNA prepared by [http://tropicalis.yale.edu/ Mustafa Khokha] from J strain frogs obtained from [http://www.urmc.rochester.edu/web/index.cfm?event=doctor.profile.show&person_id=1001617&display=for_researchers Jacques Robert], sequencing each to multiple-fold coverage of the genome.
 
 
The primary data from this project will be made available as soon as possible for use by the community. We plan to periodically post reports on our progress below.
 
  
 
= Assembled transcripts from RNA-seq data =
 
= Assembled transcripts from RNA-seq data =
Line 54: Line 38:
 
** 10,790 pub_long sequences are mapped to 5,283 v2_ref sequences.
 
** 10,790 pub_long sequences are mapped to 5,283 v2_ref sequences.
 
** 14,007 pub_long sequences are mapped to 7,977 v3_ref sequences.
 
** 14,007 pub_long sequences are mapped to 7,977 v3_ref sequences.
 +
 +
 +
= CHORI-219 BAC sequencing =
 +
We have started the first runs by sequencing 96 BACs from the [http://bacpac.chori.org/library.php?id=323 CHORI-219] library (vector: [http://www.sanger.ac.uk/Teams/Team53/psub/sequences/pbacgk.shtml pBACGK1.1]) at ~100X coverage. The selected BACs include ~70 genes of interest (Shroom3, Wnt5a, Glypican-4, Noggin, Gremlin, Pax6, Formin, etc., as initially identified by the group of [http://www.jgi.doe.gov/whoweare/cheng.html Jan-Fang Cheng] via probing the CHORI-219 library), as well as 10 BACs that have already been sequenced by the [http://www.jgi.doe.gov/ DOE Joint Genome Institute]/[http://www.hudsonalpha.org/genome-sequencing-center HudsonAlpha Genome Sequencing Center] to serve as positive controls for the sequencing and assembly pipeline. 
 +
* CHORI-219 BACs: [[xdata:BACsFor1Percent.xls| List of 96 test BACs]] (MS Excel file)
 +
 +
See [[/XENLA_SA09023]] for more details. Three mate paired libraries were sequenced:
 +
* X_laevis_WG - the ''X. laevis'' whole genome library, 5kb insert size - about 4.4GB raw data, 0.4GB high quality data<br>
 +
* X_laevis_2kb - The set of 96 BACs, with 2kb insert size - about 3.6GB raw data, 0.3GB high quality data<br>
 +
* X_laevis_5kb - The set of 96 BACs, with 5kb insert size - about 2.8GB raw data, 0.2GB high quality data<br>
 +
This (very roughly) corresponds to >600X coverage by raw data, ~50X coverage by high quality data, of the BAC set.
 +
* Given that we currently see better mapping of the shotgun SA09023 reads to ''X. tropicalis'' than to ''X. laevis'' (both to BACs and mRNAs), we're confirming the sample identity before continuing with whole genome sequencing. See the 'sanity check' [[/Species_Identification]] for details.
 +
 +
= J-strain whole genome sequencing =
 +
In addition, we are generating several mate pair libraries of different sizes from genomic DNA prepared by [http://tropicalis.yale.edu/ Mustafa Khokha] from J strain frogs obtained from [http://www.urmc.rochester.edu/web/index.cfm?event=doctor.profile.show&person_id=1001617&display=for_researchers Jacques Robert], sequencing each to multiple-fold coverage of the genome.
 +
 +
The primary data from this project will be made available as soon as possible for use by the community. We plan to periodically post reports on our progress below.
  
 
= References =
 
= References =

Revision as of 22:37, 23 October 2011

Xenopus-PV.jpg

Xenopus laevis is an essential model organism in several areas of biology. In addition to the key attributes of these embryos for in vivo imaging, cell-free extracts from Xenopus provide among the most powerful in vitro systems for studies of cell and molecular biology. A complete sequence of the X. laevis genome is an essential resource for accurate identification of peptides for mass-spec analyses, for cloning of an ORFeome, for identifying evolutionarily conserved regulatory regions, and for design of morpholino-oligonucleotides for gene knockdowns.

The Wallingford and Marcotte labs have obtained funding from the Texas Institute for Drug and Diagnostic Development (TI3D) to begin sequencing of the X. laevis genome. We are primarily working with Scott Hunicke-Smith at the University of Texas Genome Sequencing and Analysis facility, with funding sufficient for ~20x coverage of the X. laevis genome using ABI SOLiD next-generation sequencing.

Contents

Assembled transcripts from RNA-seq data

Disclaimer

  • Data users may freely download and analyze data. They may use data in publications focused around individual genes.
  • Data users may use data to analyze their own data, i.e. reference database for MS/MS proteomics data, and/or RNA-seq data.
  • The publication and presentation of global analysis of data with these sequences are not allowed until 'data owner' published the paper. As soon as the paper is accepted, we will post that info on this website.


If you have any question about this data in general, please contact to Taejoon Kwon.

TXGP201107_XENLA_EGG

Contact info:Edward Marcotte,Taejoon Kwon.

Park201106_XENLA

(Courtesy of Tae Joo Park & Richard Harland, University of California at Berkeley) Contact info: Tae Joo Park, Richard Harland.


CHORI-219 BAC sequencing

We have started the first runs by sequencing 96 BACs from the CHORI-219 library (vector: pBACGK1.1) at ~100X coverage. The selected BACs include ~70 genes of interest (Shroom3, Wnt5a, Glypican-4, Noggin, Gremlin, Pax6, Formin, etc., as initially identified by the group of Jan-Fang Cheng via probing the CHORI-219 library), as well as 10 BACs that have already been sequenced by the DOE Joint Genome Institute/HudsonAlpha Genome Sequencing Center to serve as positive controls for the sequencing and assembly pipeline.

See /XENLA_SA09023 for more details. Three mate paired libraries were sequenced:

  • X_laevis_WG - the X. laevis whole genome library, 5kb insert size - about 4.4GB raw data, 0.4GB high quality data
  • X_laevis_2kb - The set of 96 BACs, with 2kb insert size - about 3.6GB raw data, 0.3GB high quality data
  • X_laevis_5kb - The set of 96 BACs, with 5kb insert size - about 2.8GB raw data, 0.2GB high quality data

This (very roughly) corresponds to >600X coverage by raw data, ~50X coverage by high quality data, of the BAC set.

  • Given that we currently see better mapping of the shotgun SA09023 reads to X. tropicalis than to X. laevis (both to BACs and mRNAs), we're confirming the sample identity before continuing with whole genome sequencing. See the 'sanity check' /Species_Identification for details.

J-strain whole genome sequencing

In addition, we are generating several mate pair libraries of different sizes from genomic DNA prepared by Mustafa Khokha from J strain frogs obtained from Jacques Robert, sequencing each to multiple-fold coverage of the genome.

The primary data from this project will be made available as soon as possible for use by the community. We plan to periodically post reports on our progress below.

References

Protocol