Experiment: Transform HB101 Strain with pGlo plasmid using Ultrasound device

Equipment:   Off the shelf Ultrasonic Jewellery cleaning box with about 1l bath.
              Strains, DNA and reagent in Dark Glass flat bottom tubes 5ml with Plastic Caps

Sonicator Essentials:
        42 KHz
        35 watt Peak
        20cm sq transducer area in center of bath
                      Bath filled with Tap water at room temperature
Transform Tubes:
    Four Tranform tubes with

    500ul of  O.N. culture of HB101 strain
    5ul of pGlo plasmid prepared thru miniprep
    1ml of 75mM CaCl2
    Two tubes with Al foil discs (two none)


    Two Control Tubes

       Same as above , but no plasmid, no Al
       Same as Transform, with plasmid and Al


Procedure:
      Each Tube was sonicated for 10 secs at only available setting on Sonicator (on/off)
      except 2nd control tube with plasmid and Al

         Test transform Tube 1 with Al placed in middle of Transducer bump, flat glass  bottom down
         Test transform Tube 2 without Al placed in middle of Transducer bump same way
         Test transform  Tube 3 with Al placed in a side of the bath away from transducer
         Test transform  Tube 4 without Al placed in a side of the bath away from transducer
         Control  Tube 1 without plasmid and Al placed in middle of transducer bump
         Control Tube 2 with plasmid and Al left alone

   
    Post sonication
        Add 1 ml LB solution
        incubate at 37C 1 hr on a bath-shaker
                      Plate on LB and Amp agar after hour and incubate for a day
                      Check plates for colony in UV light next day



Results:
    Test tube

    With HB/PGlo/Al/midTransducer: 9 Colonies
    with HB/PGlo/midtransducer: 4 colonies
    with HB /PGlo/Al/side placed: 3 colonies
    with HB/Pglo/side placed: 1 colony


     Control
    1. Just bacteria/midplaced: 0 colony
           2. Bacteria/Pglo/Al no sonication: 0 colony

FWIW, we are pretty happy about it...open to all ideas to improve it.