Download - Buenos Dias!
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Buenos Dias! Meeting time for next week? Undergrad Research Symposium Reminder – meeting with professors Friday the 16th
from 5-6:30. Movie Night on Thursday
Real Genius/Caprica – other suggestions? New Location Safety Exams!? Safety Meetings on the 10th and 17th
9-12 or 10-1?? Check out these two Generally Informative Syn Bio
Documents. Nice Job at EOH (Pictures!) Projects can be classified as dealing with a specific
component of a system: inputs, information processing, or outputs.
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General Lab Basics
• While in lab remember act professional– Just be courteous
• Keep a great lab notebook– Always write everything down every time
• All information will be in duplicates (triplicates with the wiki)– One for the in-lab notebook; One for your
notebook; (And one for the wiki)• Try to keep lab space as clean as
possible– Dishwashing, autoclaving and disinfecting will
be covered a Saturday
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Bacterial Growth
•About 2-4 hours Log phase begins•8-10 Stationary phase begins•12 hours = Overnight (middle of Stationary phase)•24+ hours es no bueno…
• We use LB liquid and solid media to grow cells• 10 grams Tryptone• 5-10 grams NaCl• 5 grams Yeast Extract• 15 grams agar (solid media)• all in a 1L dH2O batch
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Cloning
Biobricks http://ginkgobioworks.com/support/
Digest Run a gel
Ligate Test resistance
From the ‘some PCR
Design primers
Ligate Test resistance
Analyze
<--TRANSFORM
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Cloning
Digestions/Ligations
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Cloning
• Transformation– Need competent cells– Done by either:
• Heat Shock Transformation using CaCl,– Wash cells with CaCl solution reagent– Relatively inexpensive
• High Efficiency Electroporation– Shorter Procedure– Need expensive cuvettes– Extremely efficient– Wash cells with 10% Glycerol
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Cloning
Gel Electrophoresis ALWAYS USE
LADDERS!!!!
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References
Current Protocols in Molecular Biology Search in Pubmed
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Stuff
Plate Reader basic info about designing primers how a pcr works. Stuff about mini/midi preps. Also, how to streak a plate, how much culture
and broth to use for a liquid suspension. How long is stuff good for in the 4deg. room.
what do you keep in the -20 vs. -80 and cryostocking stuff.
How to do ligations and digestions When to autoclave pHing stuff – when and how Nano-drop (it like it’s hot) List of Supplies
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Questions!?
What procedure type stuff did we forget? What do you guys know and consider important? What questions do you have?
Questions about the regional conference.