Team:UCSC/Experiments

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Experiments

Molecular Cloning

▶ LB Medium and LB/Kan plates

▶ Plasmid elution and Transformation into DH5-α

▶ Index Plating

▶ Liquid Cultures and Colony PCR

▶ TBE Buffer and Gel Production

▶ Plasmid Miniprep from DH5-α

Site Directed Mutagenesis

▶ Primer production and NEB Site-Directed Mutagenesis

▶ Transformation into Zymo Mix&Go DH5-α

▶ Plasmid Miniprep from DH5-α(Refer to Molecular Cloning #4)

Protein Production and Quantification

▶ Large Scale Protein Production and Purification

▶ Transform plasmid into XJb (DE3) Mix&Go(Refer to Site-Directed Mutagenesis #2)

▶ Start Liquid Cultures of XJb (DE3)(Refer to Molecular Cloning #3)

▶ Induction of Protein Expression via IPTG

▶ Purification via Heat-Soluble Precipitation and His-Spin Column

▶ Histidine Purification through Gravity

Thermostability Testing

▶ Incubation of Embryonated Chicken Eggs and Candling Procedure

▶ Red Blood Cell Preparation and Hemagglutination Assay

▶ Fibroblast Cell Culture & Plaque Assay