BIOL 4340/4341 Final Exam Review
Lab 6
Know how to handle protein
Disturbing tissue
Keep pH same as cell
B-mercprto ethanol dithiothreitol *** know their structure and what they’re used for
Osmoticants- sucrose and sorbitol
Detergents- triton and SDS
Lyophilization and reverse osmosis
Hofmeister series
Denature definition
Know advantages and disadvantages between two assays: Bradford and UV-280 protein
assay
Know all the blue regeants
Coomassie brilliant blue- binds to protein and used in Bradford
Different answers for Bradford and Uv-280 because of
Measuring amounts of proteins in an aliquot
Beers law---concentration moles per liter
Make sure it was due to an aliquot of 25 ml
Lab 7
Types of chromatography
Phases
Forms
Tell the difference between gel, ion, and affinity
Stationary phase, diethylameden stationry phase, question not asked.
Main sugar and main linkage
OD620 (– – –)
(DNP-amino
OD400 (——)
(Blue Dextran)
Elution Volume / Fraction No.
know peaks
(lysozyme and BSA)
BSA is biggest in the middle
calculate the resolution between two peaks
identify peaks
Kav= Ve-Vo/Vt-Vo
Use a standard graph to find weight
Lab 8
Know hormone for pregnancy test and what ELISA we were using (we used indirect)
known sandwich and direct
Theory and components and electrophoresis
Gel matrices
Starch
Agarose
Polyacrylamide
Ammonium persulfate: generates (becomes) free radicals
Methylene-tris-acrylamide crossing agent
V= E x q/ f
Gel filtration (sample components have the opportunity to go through vs gel
electrophoresis (have to go through the pores)
Bromophenol blue- low MW and is a tracking dye helps for Rf
Tracking dyeKnown about SDS- whats it used for
10^ of the y axis to find the real MW
Known an obvious strip BSA lysozyme, large subunit, ladder
ELISA
antibodies
Lab 9
Forms of DNA that we are extracting
Heterochromatin vs euchromatin
Ethanol precipitation
Remember the UV260/280 chart
1.8- he likes 1.8
sticky ends
restriction enzymes
safety mechanism
restrict the take over of their cells by virus/ bacteria
ethidium bromide (mutagen) vs DAPI
given three or four standards and do log10 and find rf to make graph
Lab 10
Know theory of centrifugation
Calculate relative centrifugal force
Of all of the things that you use that spin and their function which one has centrifugal
force as an important part of how it works (requires)
Centrifuge and the washing machine and the decanter
Don’t have to known density but know theres a transition and know their rank
Stoke’s law
s (ratio) and S (Svedberg)
G- in terms of gravities
Know ribosome size
Differential vs density centrififugation
Which one did we use
DIFFERENTIAL (enriched fractions)
Isopycinc gradient is the best
Marker enzymes which ones we used
Succinate- mitochondria
Phosphoglucoisomerase- cytosol
Cyanide to block
Sum reaction