User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/10/16

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PCR of ADA Mutations

Goals

  • Prepare ADA primers in order to conduct a polymerase chain reaction (PCR)
  • Run HRP Luminol Assay

Procedure

  • Assigned Primer= ADA K110A F (forward primer)
    • Sequence Issue number: 87635082
    • Sequence: ATT TGG CCA AAT GGG CAG TTA TTG AA
    • TM= 65.5°C
    • MW= 16833.0 g/mol
    • 33.4 nm= .56 mg ( amount of present plasmid)

concentration of plasmid needed for experiment= 100ng/μL

.56mg= 5600 ng

M1V1= M2V2 5600 ng/μL * V1= 100ng * 1000μL

    • (The container containing the plasmid only had room for 1mL of water to be added- as a result, a dilution needed to be done in order to get the appropriate concentration.