[指南]天津医科大学专业外语2.ppt

Medical English;Improve Your Spoken English;Improve Your English Writing;Recommended Readings for English in General;Acquire Basic Knowledge;Reading a Paper;Reading a Paper;Read a Paper;Reading a Paper;Participate a Journal Club;Scientific Ideas;Case Study;Propose a Hypothesis;Test the Hypothesis;Talk to Your Mentor;A Good Example;A Bad Example;;Study the Protocol;4. Add 100ul freshly prepared lysis buffer (50ul 400mM NaOH, 50ul 2% SDS) and mix gently by inverting 5-6 times at room temperature. To make lysis buffer mix equal volumes of 800mM NaOH and 4% SDS solutions. The mixture should appear translucent and mucous-like. 5. Add 120ul solution K (neutralization buffer)(5M potassium acetate, pH5.5) and mix gently by inverting 5-6 times, incubate at room temperature for 3 min. This solution is kept in the refrigerator. The mixture should contain flocculent white precipitate at this point. 6. Remove bacterial debris by centrifugation at 12000rpm for 2 min, transfer supernatant to a fresh microcentrifuge tube. The precipitate is sticky. To transfer use a 1 ml pipet tip, depress pipet plunger, move tip to bottom of tube and aspirate supernatant. When transfering to new tube, do not touch outside of pipet tip to new tube - avoid transfer of precipitate to the new tube. ;7. Add 200ul isopropanol (2-Propanol) to precipitate plasmid DNA from supernatant. Mix thoroughly by inverting 10 times. Incubate at room temperature for 1 min. 8. Collect DNA by centrifugation at 14000 rpm for 1 min. Pour off isopropanol into beaker. 9. Add 500ul 70% ethanol to tube, mix by inverting (brief vortexing okay too). Spin the DNA down at 14000rpm (full speed) for 1min. 10. Pour off ethanol. Respin for 1 min. Use 100 ul pipet to carefully remove remaining ethanol. Air dry DNA for 10 min. 11.Dissolve DNA in 35 ul water. The final concentration of DNA should be approximately 0.5 ug/ul.;Essay