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WSSP: Duckweed

WSSP: Duckweed. Pickin ’ your colonies Overnights Blue and White colonies . The cDNA bacteria (hopefully containing piece of the duckweed genome) is growing on the plate. Note the two colors of the colonies.

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WSSP: Duckweed

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  1. WSSP: Duckweed Pickin’ your colonies Overnights Blue and White colonies

  2. The cDNA bacteria (hopefully containing piece of the duckweed genome) is growing on the plate. Note the two colors of the colonies. • The bacteria is grown on a medium with X-gal. This synthetic molecule when cleaved appears blue. • The blue colonies have the ability to break down the X-gal and turn the surrounding media blue. • The white colonies cannot break down the X-gal. • Which colonies will have the “insert” of Duckweed cDNA? B-galactosidase X-gal Galactose The “other” molecule will appear blue

  3. pTriplEX plasmid This is the Lac Z gene which make B-galactosidase. Functional B-galactosidase will digest X-gal.

  4. Makes B-galactosidase Lac Z gene What color are the colonies? PtripEX plasmid Cannot make B-galactosidase cDNA insert Lac Z gene PtripEX plasmid What color are the colonies?

  5. Cloned in cDNAfragment from duckweed. Possible 900 bases long. What is the size of the plasmid with the insert? Does this plasmid make functional B-galactosidase? What color is the colony on the plate?

  6. Small, freshly saturated cultures of E. coli are called overnights (ONs). Each person in each group will prepare one overnight. Each person should label one culture tube with the proper clone name. 17DOM1.10 Your school # your Initials, your clone number, year Write name of your clone directly on the side of the tube before you add the LB+amp. Do not write on the caps!Capscan be switched, causing strains to be mixed up. Cloudy culture indicates bacterial growth Clear culture indicates NO bacterial growth

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