Molecular Biology Problem Solver 50 ppsx

... 521 Planning the Baculovirus Experiment . . . . . . . . . . . . . . . . . . . . . 521 Molecular Biology Problem Solver: A Laboratory Guide. Edited by Alan S. Gerstein Copyright © 2001 by Wiley-Liss, ... an Appropriate Expression Host . . . . . . . . . . . . . . . 501 Selecting an Appropriate Expression Vector . . . . . . . . . . . . . 506 Implementing the Eukaryotic Expression Expe...

Ngày tải lên: 02/07/2014, 04:21

... expression, 502 503 Transilluminators, crosslinking on, 422 Translation, with prokaryotic promoters, 464 Translation termination, problems with, 483–484 Trichoplusia ni, in eukaryotic expression, 503 , ... 349 electrical power for, 350 353 native PAGE versus, 345–348 problems with, 480–481 standardized gels for, 363 Sealed containment rooms, in biosafety, 117 Secondary antibodies, 384 prob...

Ngày tải lên: 02/07/2014, 04:21

... . . . . . . . . . . 150 A Wipe Test Detected Radioactivity on the Outside of the Vial. Does This Indicate a Problem? . . . . . . . . . . . . 151 Molecular Biology Problem Solver: A Laboratory ... Reference Date Has Long Passed? . . . . . . . . . . . . . . 150 Handling Radioactive Shipments . . . . . . . . . . . . . . . . . . . . . . . . . 150 What Should You Do with the Radioactiv...

Ngày tải lên: 02/07/2014, 04:21

... to the reaction, but might not be 146 Volny Jr. Becquerels, divide by 27 (27.027): 50mCi = 50 ¥ 10 -6 Ci = (50 ¥ 10 -6 Ci ¥ 3.7 ¥ 10 10 dps/Ci) = 1.85 ¥ 10 6 dps = 1.85 ¥ 10 6 Bq = 1.85 MBq.) where A 0 is ... 1983). At a specific activity of 3000 Ci/mmol, that 50 mCi translates to 16.6 femtomoles of 32 P dNTP being added to the reaction mix, while 50mCi of a 32 P labeled dNTP at a speci...

Ngày tải lên: 02/07/2014, 04:21

... less than 50% . Since recoveries will vary with sample origin, consider making your plans based on yields pub- lished for samples similar if not identical to your own. Do You Require High Molecular ... DNA and subsequent ethanol washes eliminate such a contaminant. Phenol can also be problematic. If you experience problems with DNA purified by a phenol-based strategy, apply chloroform to ex...

Ngày tải lên: 02/07/2014, 04:21

... the RNA with a low speed spin, (approximately 3000 ¥ g; approximately 7500 rpm in a microcentrifuge, or approximately 5500 rpm in a SS34 rotor), for 5 to 10 minutes at room temperature or at ... 15 minutes 50 ml 12,000 ¥ g 15 minutes How Many Rounds of Oligo(dT)–Cellulose Purification Are Required? One round of poly(A) RNA selection via oligo(dT)–cellulose typically removes 50 to 70% ....

Ngày tải lên: 02/07/2014, 04:21

... Processivity Extension (3¢–5¢ 5¢–3¢ 50% activity (dNTP/ Rate (dNTP/ Enzyme exonuclease) Exonuclease remains) binding) s/mol) Taq DNA Absent Present 9 at 97.5°C 50 60 60– 150 polymerase (40–60 at 95°C. ... Absent Present 150 at 96°C polymerase (Dynazyme TM ) e Platinum Pfx f Present Absent 720 at 95°C 100–200 100–300 180 at 100°C Platinum Taq f Absent Present 96 at 95°C 50 60 60– 150 A...

Ngày tải lên: 02/07/2014, 04:21

... recombinant adenoviruses. Proc. Nat. Acad. Sci. U.S.A. 95: 2509 –2514. Higgins, D. R., and Cregg, J. M. 1998. Pichia Protocols: Methods in Molecular Biology, vol 103. Humana Press, Totowa NJ. Hill-Perkins, ... to 50 passages. At that time a fresh culture is started from frozen cells. IMPLEMENTING THE BACULOVIRUS EXPERIMENT What’s the Best Approach to Scale-Up? Cells are initially grown i...

Ngày tải lên: 02/07/2014, 04:21

... microbiology labs, 120 pI pH gradients and, 366–368 2-D gels for determining, 365–366 Pichia methanolica, eukaryotic expression with, 505 506 Pichia pastoris, eukaryotic expression with, 505 506 Pipettes, ... 411 Probe templates, in hybridization experiments, 402 Problem reduction, 2 Problem resolution, failed, 28 Problems. See also Global problems; Technical problems with acrylamide...

Ngày tải lên: 02/07/2014, 04:21