Team:NTNU-Trondheim/Experiments

Experiments

Protocols for all the various techniques and procedures are presented in detail in this section. The protocols are divided into three parts, one set of protocols for the heme approach, one set for the sulfide-responsive transcriptional repressor and protocols used by both approaches.

Heme Protein Approach

• Preparing potassium phosphate buffer (PPB) and stock solutions
• Detecting (H₂S) with labeled myoglobin

Sulfide-Responsive Transcriptional Repressor Approach

• Sulfide and GSSH solution
• Sulfide solution for direct contact with bacteria

• Polymerase chain reaction
• Gel analysis
• Dpn1 enzyme digest
• PCR purification
• Gibson assembly
• Heat shock transformation
• Isolation of plasmid DNA
• Enzyme restriction and gel analysis
• Sanger sequencing

Protocols used by both approaches

Methylene blue method

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