Protocols for methods used during previous GBE courses

Click on the link for copy of the protocol and a list of required materials/reagents

General microbiology           

           Gram Staining

           Aseptic technique for plating bacteria

           Serial dilutions

           Overnight cultures (from transformed colonies)

DNA extraction

            Qiagen DNEasy Method** – Kit method, suitable for most samples, long term storage possible

            Gentra DNA isolation** – suitable for isolating bacteria from cultures, insects and other environmental sources

            Squish Buffer Extraction – suitable for extracting DNA from small invertebrates; limited shelf life

Cloning

            Promega pGEM-T Easy**

            Invitrogen TOPO-TA**

            Eppendorf MINI-prep** – isolation of plasmids for sequencing

            Alkaline lysis – isolation of plasmids for sequencing

PCR

            PCR from gDNA

            Colony PCR to verify cloning

            Gel Electrophoresis

PCR Purification
            QIAquick PCR purification** - kit-based PCR cleanup for cloning

            Exo-Cip DNA Cleanup – simple 2 enzyme method to prepare samples for sequencing

Recipes for reagents and supplies