Team:Cambridge/Labwork/Protocols
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- | A list of all protocols developed during the project. | + | A list of all protocols developed during the project. We used this page as a reference throughout the competition. |
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'''Gel Electrophoresis by SDS PAGE''' | '''Gel Electrophoresis by SDS PAGE''' | ||
*[[Team:Cambridge/Protocols/Gel_Electrophoresis_of_Protein | Protein Identification by SDS PAGE]] | *[[Team:Cambridge/Protocols/Gel_Electrophoresis_of_Protein | Protein Identification by SDS PAGE]] | ||
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Revision as of 20:09, 20 September 2011
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Protocols
A list of all protocols developed during the project. We used this page as a reference throughout the competition.
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.
- 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.
- Norgen Proteospin inclusion body prep. : A proprietary kit.
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