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1、Biotech Handout Biotech Fall, 06Transformation E. coli XL-1 Blue cells with Plasmid DNAMaterialsXL-1 Competent CellsPlasmid DNASOC or LB mediumLB /amp and LB plateBio-Rad Gene Pulse apparatusStep1. Set the Bio-Rad Gene Pulse apparatus to 2.5 kV, 25 F. set the pulse controller to 200 or 400 ohms.2. A
2、dd 0.1g plasmid DNA and H2O in 2 l to tubes containing 50 l thawed XL-1 competent cells (on ice). See figure below. Mix by tapping the tube or by swirling the cells with the pipettor.Add 0.1g plasmid DNA mixAdd 2l of H2O 50 l of XL-blue cells 3. Transfer the DNA and cells into a 0.2 mm cuvette that
3、has been chilled 5 min on ice, shake slightly to settle the cells to the bottom, and wipe the ice and water from the cuvette with a Kimwipe.4. Place the cuvette into the sample chamber.5. Apply the pulse by pushing the button or flipping the switch.6. Remove the cuvette. Immediately add 500 ml SOC m
4、edium and then incubate 30 to 60 min with moderate shaking at 37C.7. Plate 30l aliquots of the transformation Mix (plasmid DNA or Water) on LB plates respectively as the figure shows below8. Put the spread plate to our incubator by inverting the culture plate and leave it overnight. Record the results and explain the results.Transformation ResultTransformation Mixl XL-1 Competent Cells with Plasmid DNAl XL-1 Competent Cells with H2OPlateLB Plate with AmpicillinLB PlateLB Plate with AmpicillinLB PlateNumber of ColoniesNote: Next time, we will perform Jelly Fish Library transformation.
