Protocols
Ligation Protocol
Gel Purification Protocol (from QIAquick Gel Extraction Kit)
Gel Electrophoresis Protocol
Digestion 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
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
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