Protocols
Ligation Protocol
Determine insert to vector ratiosCalculate the amount of insert needed if 50ng of vector is used (can use different amount of vector)In a PCR tube add the following:50ng of vectorAmount of insert based on ratios (calculated in second step)2uL of buffer2uL of DNA ligaseAmount of water to bring total volume to 20uL
Incubate overnight at 14oCNote: We used T4 DNA ligase and buffer from NEB
Gel Purification Protocol (from QIAquick Gel Extraction Kit)
Gel Electrophoresis Protocol
Preparing LB+Appropriate Antibiotic Protocol
Preparing Agar Plates Protocol (Makes 12 (15mm) Plates)
Preparing Competent Cells Protocol
Miniprep Protocol (from QIAprep Spin Miniprep Kit)
Preparing Glycerol Stock Protocol
Transformation Protocol
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