Molecular Biology Problem Solver 50 ppsx
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...
Molecular Biology Problem Solver 58 ppsx
... 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...
Molecular Biology Problem Solver 15 ppsx
... . . . . . . . . . . 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...
Molecular Biology Problem Solver 16 ppsx
... 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...
Molecular Biology Problem Solver 18 ppsx
... 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...
Molecular Biology Problem Solver 22 ppsx
... 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% ....
Molecular Biology Problem Solver 31 ppsx
... 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...
Molecular Biology Problem Solver 54 ppsx
... 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...
Molecular Biology Problem Solver 57 ppsx
... 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...
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