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P r o t o c o l s

Protocols

Find all our experimental protocols in PDF format here.

Find our protocols contribution for iGEM in the hydrogel category

Calculations

Dilution calculation

Detailed method for calculating necessary dilutions in the laboratory. This protocol includes formulas and practical examples for preparing solutions at specific concentrations.

Microbiology

Medium preparation

Protocols to prepare specialized bacterial culture media. Includes DL-glutamic acid supplementation for auxotrophic strains, tyrosine plates for melanin detection, and M9 minimal media formulations with microwave sterilization methods.

Molecular Biology

Q5 PCR

High fidelity PCR amplification protocol using Q5 enzyme. Optimal conditions for specific and efficient amplification of DNA fragments.

Molecular Biology

Colony PCR Econotaq

Rapid screening method for bacterial colonies by PCR using Econotaq enzyme.

Molecular Biology

Supercompetent cell transformation

Protocol for transforming supercompetent bacterial cells with plasmid DNA. Method for obtaining high transformation efficiency.

Analysis

Electrophoresis

Complete protocol for nucleic acid separation by agarose gel electrophoresis.

Microbiology

Supercompetent cell preparation

Method for preparing supercompetent bacterial cells for transformation. Detailed protocol to obtain high transformation efficiency.

Microbiology

Electrocompetent cell preparation

Protocol for preparing electrocompetent bacterial cells for transformation by electroporation. Method for high transformation efficiency.

Molecular Biology

Clean up

Protocol for purification of PCR amplification products and enzymatic digestions. Purification methods by column and precipitation.

Molecular Biology

Miniprep

Small-scale plasmid DNA extraction protocol. Method for isolating high quality plasmid DNA from bacterial cultures.

Molecular Biology

Golden Gate

Golden Gate assembly protocol for vector construction. Standardized assembly method using type IIS restriction enzymes.

Molecular Biology

SLIC (Sequence and Ligation Independent Cloning)

SLIC cloning protocol for assembling DNA fragments without ligation. In vitro homologous recombination method for vector construction.

Molecular Biology

Enzymatic digestion

DNA digestion protocol with restriction enzymes. Optimal conditions for complete and specific digestion of plasmid or genomic DNA.

Analysis

Cell growth measurement: OD 600

Protocol for measuring optical density at 600 nm to assess bacterial growth. Standard method for monitoring bacterial culture growth.

Microbiology

Chromosomal deletion λ-Red system

Specific protocol for construction and analysis of murI mutants. Methods for inactivation of the chromosomal murI gene and resulting phenotypic analysis.

Analysis

Western blot

Complete Western blot protocol for detection of proteins.

Hydrogel

Alginate hydrogel synthesis & bacterial encapsulation

Our contribution to iGEM is this complete protocol for hydrogel preparation using alginate and protocol for bacterial encapsulation.

Hydrogel

Hydrogel bacterial escapte test

Our contribution to iGEM is detailed protocol for testing the escape of bacteria in hydrogel

Hydrogel

Hydrogel bacterial survival test

Our contribution to iGEM is complete protocol for testing the survival of bacteria in hydrogel.

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