The Tough Tunicate - Lab Notebook

Alkaline Comet Assay For Epithelial Cells of Botryllus schlosseri

Alkaline Comet Assay: Denatures DNA at a high pH (typically around 13), which allows it to detect not only double-strand breaks (DSBs) but also single-strand breaks (SSBs)…

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Nickel x Botryllus schlosseri -omics

Field sourced Botryllus schlosseri are brough from the City of De Moines Marina on a weekly basis. Botryllus schlosseri colonies are brought back from the field and kept in…

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December Goals

goals

Update my CV with new teaching stuff and such and the conferences I presented at this year.

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SOD assay for nickel LC50 samples

An LC50 determination for nickel on Botryllus schlosseri was completed in Spring Quarter 2024. Specimens were snap frozen at the 24 and 96 hour mark to assess for…

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Nickel LC50 Experiment 3

study plans

nickel chloride

Review resources prior to handling nickel chloride:

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Nickel LC50 Experiment 2

study plans

nickel chloride

Review resources prior to handling nickel chloride:

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Nickel LC50 Experiment 1

study plans

nickel chloride

Divided animals into separate system, 3 days prior to start of exposure, 8 days acclimated to the recirculating system.

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Troubleshoot qPCR for Nickel exposure on Botryllus schlosseri

re: RobertsLab/resources issue #1834

nickel chloride

qPCR

primers

issues

In response to @kubu4’s questions in issue #1834 opened in Roberts Lab resources repo.

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qPCR for Nickel exposure on Botryllus schlosseri

Relative standard curves to determine primer effenciencies.

nickel chloride

qPCR

primers

Analysis of gene expression with quantitative Polymerase Chain Reaction (qPCR) will be a component of the in vivo nickel exposure work conducted in autumn 2023. Four…

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Tunicate Hemolymph Extraction for Media Supplementation

epithelial

primaries

paper

Protocol replicated from following paper.

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Amino acid supplementation to media in Sea Sponge cell line.

paper

Hesp K, Van Der Heijden JME, Munroe S, Sipkema D, Martens DE, Wijffels RH, et al. First continuous marine sponge cell line established. Sci Rep. 2023 Apr 8;13(1):5766.

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Primary Epithelial Cell Culture Protocol

epithelial

protocol

This protocol is version 02.04.24

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Botryllus schlosseri Epithelial Tissue Isolation Protocol

epithelial

protocol

This protocol is version 01.25.24

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Nickel Impacts on Morphology of Botryllus schlosseri

Analysis of pilot study.

nickel chloride

Data below was collected from a pilot study conducted in Autumn 2023 where field-collected individual Botryllus schlosseri were exposed to a low and high treatment of nickel…

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Nickel Impacts on Primary Epithelial Cell Cultures of Botryllus schlosseri

Analysis of pilot study.

nickel chloride

primary cultures

epithelial

Data below was collected from a pilot study conducted in Autumn 2023 where field-collected individual Botryllus schlosseri were exposed to a low and high treatment of nickel…

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Nickel chloride x Botryllus schlosseri Exposure 3

study plans

nickel chloride

Divided animals into separate system, 3 days prior to start of exposure, 8 days acclimated to the recirculating system.

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Designing Tunicate Primers

protocol

qPCR

primers

nickel chloride

I am interested in designing primers for Botryllus schlosseri to evaluate differences in expression of apoptosis, stemness, oxidative stress, and senescence related genes…

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Exposure 2 in vivo NiCl₂ x Botryllus schlosseri

study plans

nickel chloride

Divided animals into separate system, 3 days prior to start of exposure, 8 days acclimated to the recirculating system.

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Lab Reared Botryllus schlosseri Care and Maintenance

protocol

This protocol is for weekly care of lab born Botryllus schlosseri. Field sourced tunicates are brought in during the months of May-October and allowed to release oozooids on…

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Pilot 1.2 in vivo NiCl₂ x Botryllus schlosseri Exposure

study plans

nickel chloride

Divided animals into separate system, 3 days prior to start of exposure, 8 days acclimated to the recirculating system.

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Use of Genetic Algorithm in Cell Culture Media Optimization

cool paper

optimization

Munroe, S., K. Sandoval, D. E. Martens, D. Sipkema, and S. A. Pomponi. 2019. Genetic algorithm as an optimization tool for the development of sponge cell culture media. In…

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Substrate Options for the in vitro Culture of Botryllus schlosseri Epithelial Cells

cool paper

discussion

Exploring of the physiology of the extra cellular matrix (ECM) of the marine invertebrate, ascidian, Botryllus schlosseri as to identify potential substrates for in vitro pri…

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Pilot 1.1 in vivo NiCl₂ x Botryllus schlosseri Exposure

study plans

nickel chloride

First pilot study for NiCl₂ exposures on whole field-sourced Botryllus schlosseri colonies.

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Trypsinization of Botryllus schlosseri Epithelial Cells

protocol

Trypsinization of cell cultures is a common method use in cell biology to disassociate adherent cells in vitro from the vessel in which they are being cultured for the…

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Masters Thesis Progress

thesis

study plans

This page is intended to act as a living document recording the progression of my thesis work.

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Cell Isolation Trial 18: Cultrex Substrates

epithelial

ampullae

zooids

primary buds

primaries

substrate

In this small scale experiment, resorbing zooids and take over zooids, two distinct asexual developmental stages sourced from the same Botryllus individual, were seeded…

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Cell Isolation Trial 17

epithelial

zooids

primaries

Here I both describe the procedure that occurred for epithelial tissue isolation from Botryllus schlosseri for trial 16 and maintain this document as a living log for…

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Cell Isolation Trial 16

epithelial

ampullae

zooids

primaries

Here I both describe the procedure that occurred for epithelial tissue isolation from Botryllus schlosseri for trial 16 and maintain this document as a living log for…

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First Aquatic Invertebrate Cell Line

cool paper

Here I summarize key takeaways and implications from a paper that describes the establishment of the only aquatic invertebrate cell line. The author is Eder L. Hansen.

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Example of Post With Code

code

analysis

This is an example post of the work I did for Assignment 5 for FISH 549 where I generated an exploratory and expository plot of a data set provided by the class.

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