Transformation Of Bacteria

Transformation of Bacteria
Hypothesis: If the petri dishes contain restriction plasmids, then they go forth have bacteria growth because they will resist the ampicillin.
Materials:
            Petri dish
            Pipet
             glaze spreader
            Rubbing alcohol
            Nutrient agar
            Boiling water
            Ice
            Plasmid
            2 of each LB + and menage, LB/Amp + and , and LB/Amp/X-gal + and
            LB broth
            DNA
            Incubator
             inoculate grummet
           
Procedure:
1.      Mark unrivalled aseptic 15 ml supply +. Mark another -.
2.      Use a sterile transfer pipet (Figure 1) to add 259 ul of ice cold calcium chloride to each pipework.  Place both tubings on ice.
3.      Use a sterile plastic inoculating tat to transfer one or two large colonies of  E.coli from the starter plate to the + tube.
a)      Be careful not to transfer any agar from plate along with cell mass.
b)      Immerse loop tip in calcium chloride solution and vigorously tap against wall of tube to dislodge cell mass.
4.      Immediently suspend cells in the + tube by repeatedly pipetting in and out, using the sterile pipet.  Do not make bubbles.



 Hold tube to light, and care intacty inspect to ingest that respite is homogeneous.   No visible clumps of cells should remain in the tube or be lost in the bulb of te transfer pipet.
5.      Return + tube to ice.  Transfer a sulphur mass of cells to the - tube and suspend as described in steps 3 and 4 above.
6.      Both tubes should now be on ice. Best if in ice for about 15 minutes.
7.      Use a sterile plastic inoculating loop to transfer one loopful of the plasmid solution to the + tube. 10 ul is measured when DNA solution forms a bubble across the loop opening. Immerse loopful of plasmid solution directly into cell suspension and swish loop to mix DNA.
8.      Return + DNA tube to ice and incubate both tubes on ice for 15 minutes or longer....If you want to get a full essay, order it on our website: Orderessay



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