카이스트 분자공학실험 Introduction of chemical and biomolecular engineering 결과 보고서
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- 2015.10.10
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- 2013.09
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한컴오피스
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목차
1. Experimental procedure
2. Results and discussion
1) Results
1-1) Data analysis
1-2) Caution
2) Discussion
3. Reference
본문내용
1. Experimental procedure
① Choose a single colony from a freshly streaked bacterial plate and use it to inoculate LB plus an appropriate antibiotic. Incubate the culture overnight while shaking.
② Harvest 3-5ml of bacterial culture by centrifugation at 13,000 rpm for 30 sec at RT and discard the supernatant.
③ Resuspend the pellet in 250μl of resuspension buffer, vortexing or pipetting until no clumps of the cell pellet remain.
④ Add 250μl of Lysis buffer to the resuspended cells. Close tube and gently mix by inverting the tube several times. Do not vortex and do not exceed 5 min of lysis time.
⑤ Add 350μl of neutralization buffer and gently mix by inverting the tube several times.
⑥ Centrifuge at 13,000 rpm for 10 min at 4℃. While waiting for the centrifugation, insert a column into the collection tube.
⑦ After centrifugation, transfer supernatant promptly into the column.
Cell debris, protein, and genome DNA will form a compact white pellet in the tube. Do not transfer with the white pellet.
⑧ Centrifuge at 13,000 rpm for 60 sec.
참고 자료
CBE201 Molecular Engineering Laboratory, KAIST, 2013
Kutzler MA, Weiner DB: DNA vaccines: ready for prime time? Nat Rev Genet 2008, 9:776–788.
Sardesai NY, Weiner DB: Electroporation delivery of DNA vaccines: Prospects for success. Curr Opin Immunol 2011, 23:421–429.
M. Chattergoon, J. Boyer, D. Weiner, Genetic immunization: a new era in vaccines and immune therapeutics, FASEB, 11 (1997), pp. 753–763
Gel Electrophoresis of DNA
Agarose Gel Electrophoresis