Protocols

A list of all protocols developed during the project. To add one, use the section below.

Amplification of DNA

• Polymerase Chain Reaction : A method for amplifying a section of DNA.
• Colony PCR : PCR with cells as a template. Useful for checking the length of an insert in an introduced plasmid.

Analysis of DNA

• Gel Electrophoresis : A method used to separate DNA fragments of different sizes.
• Gel Extraction of DNA : A technique used to isolate a desired fragment of intact DNA from an agarose gel following agarose gel electrophoresis.
• Rescue Precipitation of DNA : Creating a clean DNA solution after dissolving agarose gel.
• Restriction Enzyme Digestion : A method for creating a restriction map of a plasmid.

Preparation of DNA Constructs

• Primer Design : Some general guidelines on how to design successful primers.
• Gibson Assembly : An extremely powerful technique for joining multiple, arbitrary DNA sequences in one step, compatible with standard assembly.

Transformation of Bacterial Cells

• Making Competent Cells : The methods required to make various cells competent
• Transformation of E.Coli : A simple method of transforming competent E.coli with your DNA of choice
• Tranformation of B. subtilis : A technique used to introduce foreign DNA into Bacillus cells.
• Transformation of E.coli by Electroporation: A technique for rapidly inserting DNA into cells, typically plasmid DNA, providing the cells are made competent in the correct manner.

Bacterial Cultures

• E. coli cell culture : A method for growing a cell culture in liquid medium
• Glycerol Stocks: A method of storing E.coli cells preserving their viability

Extraction of DNA

• Mini Prep : Extracting DNA from bacterial cells
• Extraction of genomic DNA from squid : Two methods to extract genomic DNA from squid tissue
• Extraction of DNA from Filter Paper  : Extraction of DNA from filter paper which is a safe way of shipping DNA

Microscopy

• Confocal Microscopy : A method to visualise reflectins from squid samples.
• Slide Preparation for Confocal Microscopy : A method to perpare slides with agarose supplement to form microcolonies.

1. Trypsinisation : A method of dispersing cells from tissue, for microscopy.

Protein Purification

• Buffer Preparation : Methods to prepare the various buffers used to purify his-tagged reflectin from inclusion bodies in E. coli.
• Inclusion Body Prep : Isolation of insoluble inclusion bodies of recombinant proteins.
• His-Trap Protein Purification : A method to purify reflectin from E. coli lysate using an affinity column.
• Acetone Precipitation of proteins : A method to concentrate solutions of protein.
• Ethanol Precipitation of proteins : A method to concentrate solutions of protein.

Thin Film Preparation

• Substrate Preparation:How to prepare a substrate for flow coating and spin coating.
• Spin Coating to Make a Thin Film: Our Spin Coating Protocol
• Flow Coating: How to flow coat a thin film.

Gel Electrophoresis by SDS PAGE

• Protein Identification by SDS PAGE

Adding new Protocols

To add a new protocol, enter the name in the box below and click new to create the new page. You must then return to this page in order to add in a link to the page by copying the code below.

Add a new link on this page with

#[[Team:Cambridge/Protocols/PROTOCOL_NAME_HERE |PROTOCOL NAME HERE]] : Insert description here