Equipment: CentrifugeMicrowave ovenGlovesElectrophoresis Materials Light table and cameraReagents:Ice bucket with icepUC19 vector DNAopdpcr productRestriction enzymes and NEB buffer: BamHI and NEB Buffer 3CIP1kb DNA ladder Autoclaved MilliQ H2OAgarose powder6x Loading Dye1x TAE/TBE Buffer
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MethodsThree test tubes of 1.5ml tubes where labelled in the start of the experiment such as “Insert Digest,” “Vector Digest,” and “(-) control” as well as the group number. We then had to add the reagents described in the table (page 61 in lab manual) in the listed order. The entire tube was flicked and quick spin for about three seconds in the microcentrifuge before an hour of incubating at 37 degrees Celsius. After incubation, the tubes must be placed on ice while waiting to be run on gel to keep it safe at all time.For us to prepare for electrophoresis 2.8g of agarose powder was added to a 125ml flask then add40ml 1x TAE/TBE Buffer (TAE: Tris Acetic acid EDTA; TBE: Tris Boric acid EDTA). The mixture was then heated in the microwave for about 45 seconds, while being vigilant for any liquid boil over and taking proper safety precautions upon removing the flask. Shaking the mixture at room temperature for 10 minutes is done to allow cooling before ethidium bromide addition (this activity was performed by the lab instructor). We noted that the ethidium bromide
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