Overview

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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

Maintaining the axolotls

Feeding

Artemia (Brine shrimp eggs)

Brine shrimp eggs are metabolically inactive and can remain in total stasis for two years while in dry oxygen-free conditions, even at temperatures below freezing. This characteristic is called cryptobiosis meaning "hidden life" (also called diapause). While in cryptobiosis, brine shrimp eggs can survive temperatures of liquid air (−190 °C or −310.0 °F) and a small percentage can survive above boiling temperature (105 °C or 221 °F) for up to two hours.

Once placed in brine (salt) water, the cyst-like eggs hatch within a few hours. The nauplii, or larvae, are less than 0.5 mm in length when they first hatch. Brine shrimp have a biological life cycle of one year, during which they grow to a mature length of around one centimeter on average. This short life span, along with other characteristics such as their ability to remain dormant for long periods, has made them invaluable in scientific research, including space experiments. This ability has also enabled the use of a hybrid of brine shrimp, bred to grow larger and live longer, as Sea-Monkeys.

Artemia Hatching Preparation: Protocol for 15 Ltr.

Switch on the bubbler.
Take the warm water from the tap and fill it into the container, meanwhile add 325 ml volume of salt (Red Sea Coral pro Salt), when the 15 Ltr. container is filled up the salt should be dissolved already.
Switch on the heating. And make sure that the light on the top is on ☺ Add 50-60 ml full of dried artemia (Stored at 40C) to this.

Hatching preparation
  • The following protocol is considered for 15 l solution.
  • Switch on the bubbler.
  • Take the warm water from the tap and fill it into the container, meanwhile add 325 ml volume of salt (Red Sea Coral pro Salt), when the 15 Ltr. container is filled up the salt should be dissolved already.
  • Switch on the heating. And make sure that the light on the top is on.
  • Add 50-60 ml full of dried artemia (Stored at 40C) to this.

Keep it like this for 2 days with aeration. And >200 Lux of light.
Usually they hatch within few hours but It is good to keep for 2 days to achieve a good hatching.
During these two days temperature can be 18°-20°C.
Make sure hatching container is not getting clogged during these two days.

Lab duties


Protocols


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.

Transgenic lines

Axolotl (A.mexicanum)

Name Construct Method of transgenesis Author Tissue Image(s) Remarks
PM28 Prrx1-TFP-NLS-T2A-mERt-Cre-ERt Sce1 Prayag Murawala Limb bud and limb blastema Germ-line transmitted
PM33 Col1A2-TFP-NLS-T2A-mERt-Cre-ERt Sce1 Prayag Murawala Bone and fibroblast Germ-line transmitted

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.

Theses

Title Summary Author Link
Test Test John Doe tba

Databases

  • 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.
  • Axologle database contains several different Axolotl transciptome assemblies

Datasets

Next generation sequencing

The data can be found under //biodata/groups/crtd_tanaka/axolotl/Sequencing/Illumina.

  • Mature and regenerating spinal cord
    Name Description Type Reads Location Author Remarks
    Mature spinal cord Mature axolotl spinal cord Illumina, paired-end, 100bp 65,753,941 SpinalCord/mSC.fastq.gz Akira Tazaki
    Injured spinal cord Injured axolotl spinal cord, 6dpi Illumina, paired-end, 100bp 78,459,950 SpinalCord/6diSC.fastq.gz Akira Tazaki
  • Axolotl limb
    Name Description Type Reads Location Author Remarks
    Mature limb Mature axolotl forelimb Illumina, single-end, 76bp 65,934,228 Limb/Mature.fastq.gz Dunja Knapp
    Amputated limb (36h) Amputated axolotl forelimb (36hpa) Illumina, single-end, 76bp 65,250,768 Limb/36h_amp.fastq.gz Dunja Knapp
    Amputated limb (72h) Amputated axolotl forelimb (72hpa) Illumina, single-end, 76bp 62,970,898 Limb/72h_amp.fastq.gz Dunja Knapp
    Lateral wound (36h) Injured (lateral wound) axolotl forelimb (36hpw) Illumina, single-end, 76bp 66,361,470 Limb/36h_lw.fastq.gz Dunja Knapp
    Lateral wound (72h) Injured (lateral wound) axolotl forelimb (72hpw) Illumina, single-end, 76bp 64,133,834 Limb/72h_lw.fastq.gz Dunja Knapp