HELP WITH SCIENCE FAIR

Sean Tabacsko hockeyst at cris.com
Sat Mar 15 13:02:18 EST 1997


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Hello ,

      I am working with clay as a transgenic substrate.I am testing to see
if clay can retain plasmid DNA for later transfer to other recipient
bacteria. I have concluded that the clay holds and transfers the plasmid
DNA to the bacteria. I am using ampicillin resistance as an indicator for
the experiment. I killed some E.coli MM 294 ampicillin resistant bacteria
in clay in an oven at 130 degrees C mixed it with sterile LB broth and
streaked some of this compound on an LB/Ampicillin plate to make sure all
of the bacteria had died. Nothing grew. I used more of the LB broth, dead
ampicillin resistant bacteria,and clay compound but this time added a non
resistant MM 294 cell suspension mid log to the mix and set on ice for 20
minutes and streked them on LB/ampicillin. There was growth which means
there was a transfer of plasmid DNA to the non resistant bacteria that
made them resistant and able to grow on the LB/ampicillin plate. I also
checked that the mid log was still non resistant before adding it to the
compound. I guess the questions I have are what sort of things do DNA like
to bind to and why did the clay act as a good substrate. I would also
appreciate any other thoughts or information that you can give to me
conserning this project or substrates. I would like any help you can give
me.



                                                                                            
Thanks in advance,



Sean A. Tabacsko
Center For The Arts And Sciences
Saginaw, MI



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