Difference between revisions of "Overview"

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<li>[[Metamorphosis_protocol|Metamorphosis protocol (Sherry)]]</li>
 
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<li>[[Digestion_DNA|Digestion (Plasmid DNA)]]</li>
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<li>[[Digestion_DNA|DNA Digestion (Plasmid)]]</li>
<li>[[Extraction_of_RNA_from_axolotl_tissue|RNA Extraction (axolotl tissue)]]</li>
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<li>[[RNA_Extraction_from_axolotl_tissue|RNA Extraction (axolotl tissue)]]</li>
  
 
== Lab business ==
 
== Lab business ==

Revision as of 12:27, 6 March 2014

Introduction

On the pages listed below you can find most common protocols used in the lab as well as a list of lab duties along with the names of the responsible persons. Please, contact the responsible persons if you have any question, suggestions or troubles.

Axolotl

Maintainance

In this section you will find some general information on how to maintain the animals.

Feeding

This section will give you an overview of how to feed the axolotls.

Interesting observations

In the following section you can find some interesting observations concerning the axolotl, its behavior, caveats of the transgenics and many more.

Lab duties


Protocols

    Cell culture

  • Blastema cells
  • Baculovirus titration
  • Myoblasts electroporation
  • Adhesive cell culture
  • Electroporation of blastema cells
  • Electroporation of neural epithelia cells (axolotl spinal cord)
  • mESC culture/Cyst formation
  • Neurosphere culture
  • Histology

  • Double Immunostaining via antigen retrieval (Sherry)
  • Molecular biology

    • Metamorphosis protocol (Sherry)
    • DNA Digestion (Plasmid)
    • RNA Extraction (axolotl tissue)
    • Lab business

      Ordering database

      Use the following ordering database to place your orders.

      Booking calendars

      Since some pieces of hardware are used intensively it is necessary to book them in advance. The links to the corresponding booking calendars are listed below.

      Genetics

      Transgenic lines

      Markers

      Marker Full name Target Remarks Link
      DAPI/Hoechst 4',6-diamidino-2-phenylindole Nuclei Incorporates into the DNA DAPI, Hoechst
      PCNA Proliferating-Cell-Nuclear-Antigen Dividing cells PCNA
      SOX2 SRY (sex determining region Y)-box 2 Preferentially neuronal lineage Progenitor marker for both spinal cord and brain SOX2
      PAX6 Paired box protein Pax-6 Brain/spinal cord aka aniridia type II protein (AN2) or oculorhombin PAX6
      PAX7 Paired box protein Pax-7 Satellite cells/spinal cord/brain PAX7
      MHC Myosin heavy chain Muscle fibers MHC
      MYF5 Myogenic factor 5 Myoblasts Regulates muscle differentiation, labels cycling myoblasts MYF5
      TUBB3 ßIII-tubulin Axons TUBB3
      MBP Myelin basic protein Schwann cells
      PRRX1 Paired mesoderm homeobox protein 1 Connective tissue progenitors in Limb bud and Limb Blastema PRRX1
      SMA Smooth muscle actin Blood vessels/endothelial cells
      GFAP Glial fibrillary acidic protein Glia GFAP
      NeuN Feminizing Locus on X-3, Fox-3, or Hexaribonucleotide Binding Protein-3 Differentiated nerve NeuN
      LEU7/HNK1/B3GAT1/CD57 Galactosylgalactosylxylosylprotein 3-beta-glucuronosyltransferase 1 Schwann cells B3GAT1
      XBRA Xbrachyury Mesoderm during gastrula stage Brachyury
      MSX1 Muscle segment homeobox, Msh homeobox 1 Undifferentiated cells Transcriptional repressor during embryogenesis MSX1
      Trypan blue Dead tissues or cells Vital stain Trypan blue
      MEF2C Myocyte-specific enhancer factor 2C Differentiated muscle cells aka MADS box transcription enhancer factor 2, polypeptide C MEF2C
      PH3 Phospho-Histone H3 Mitotic cells
      BrdU Bromodeoxyuridine Cells in S-phase BrdU
      CASP3 Caspase 3 Dying cells CASP3
      GDF5 Growth/differentiation factor 5 Joints Joint development marker GDF5
      Survivin Cleavage furrow aka baculoviral inhibitor of apoptosis repeat-containing 5 or BIRC5 Survivin
      MAP2 Microtubule-associated protein 2 Neurons MAP2
      SOX9 SRY (sex determining region Y)-box 2 Cartillage progenitors SOX9
      MBP Myelin basic protein Myelin sheath MBP

      Digital data

      Over the years the members of the lab have produced a lot of different data: Sanger sequences, EST libraries, images and so on. Select a category from the list below in order to view or download the data.

      Fileservers

      Location Volume size Description Access Location (console only)
      //biodata/groups/crtd_tanaka 16 TB General purpose storage space Entire Tanaka lab biocluster: /group/crtd_tanaka
      //biodata/groups/tanaka-ests 247 GB EST sequences Entire Tanaka lab biocluster: /group/tanaka-ests
      //biodata/groups/axolotl-transcriptome 500 GB Axolotl transcriptome data Elly, Sergej biocluster: /projects/axolotl-transcriptome
      //biodata/groups/tanaka-saori 878 GB Special directory for Saori's data Elly, Saori, Sergej biocluster: /group/tanaka-saori

      Tip.png NOTE: The path is different, when accessing the file server from the console.

      Literature

      Databases

      • Axolotl transcriptome website contains the Axolotl transcriptome assemblies
      • Newt transcriptome website contains the Newt transcriptome assembly
      • Red spotted newt transcriptome website contains the red spotted newt transcriptome assembly
      • Axolotl EST database contains several ESTs and also some additional information, e.g. the position of the well with the corresponding insert in the Blastema (BL) or Neural tube (NT) library.
      • Short insert cDNA library
      • CURRENTLY UNAVAILABLE! Axologle database contains expression profiling data of Axolotl limb regeneration
      • CURRENTLY UNAVAILABLE! Axolotl BLAST database contains several different BLAST-able Axolotl transciptome assemblies

      Datasets

      Interesting links

      Lab life

      • Lab photos
      • Further pictures can be found in /biodata/groups/crtd_tanaka/LabFun/
      • The recepies for Heino's farewell present can be found under /biodata/groups/crtd_tanaka/LabFun/Heino's farewell present/
      • The video of the visit of the EU commissioner can be found under /biodata/groups/crtd_tanaka/LabFun/