Sulfolobus acidocaldarius

EdU staining protocol for Sulofolobus

This protocol describes how to stain EdU after its incorporation into Sulfolobus acidocaldarius. EdU incorporation allows visualization of newly synthesized DNA, making it a useful method for studying the cell cycle. The staining process involves an EdU Click-It reaction using Alexa Fluor azide and subsequent washing steps before analysis by microscopy or flow cytometry. Compared to other DNA labeling techniques, this protocol offers high specificity and ease of implementation.

EdU Staining Protocol for Sulfolobus acidocaldarius

This protocol describes how to stain EdU after its incorporation into Sulfolobus acidocaldarius. EdU incorporation allows visualization of newly synthesized DNA, making it a useful method for studying the cell cycle. The staining process involves an EdU Click-It reaction using Alexa Fluor azide and subsequent washing steps before analysis by microscopy or flow cytometry. Compared to other DNA labeling techniques, this protocol offers high specificity and ease of implementation.

Protein Extraction and Western Blotting for Sulfolobus

This protocol describes the step-by-step methodology for extracting proteins from cells and analysing them via Western blot. The workflow includes cell lysis, protein quantification, SDS-PAGE electrophoresis, protein transfer to nitrocellulose, antibody incubation, and visualisation using Licor technology. The protocol ensures reproducible results and is adaptable for whole proteome analysis. Alternative lysis buffers, such as RIPA or TK150, may be used depending on downstream applications.

Ethanol Fixation Protocol for Sulfolobus acidocaldarius

This protocol describes the ethanol fixation of Sulfolobus acidocaldarius, which is used for immunofluorescence and flow cytometry applications. Ethanol fixation is a widely used method to preserve cell morphology and antigenicity while ensuring long-term sample storage. Alternative fixation methods include methanol and formaldehyde fixation, which offer different advantages depending on the downstream application. This protocol details the step-by-step preparation of ethanol-fixed samples, ensuring optimal preservation for cytometric and imaging analyses.

Protocol

ChIP-Seq Occupancy Mapping of the Archaeal Transcription Machinery

Genome-wide occupancy studies for RNA polymerases and their basal transcription factors deliver information
about transcription dynamics and the recruitment of transcription elongation and termination
factors in eukaryotes and prokaryotes. The primary method to determine genome-wide occupancies is
chromatin immunoprecipitation combined with deep sequencing (ChIP-seq). Archaea possess a transcription
machinery that is evolutionarily closer related to its eukaryotic counterpart but it operates in a

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