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.

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

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

Cloning

Biobricks http://ginkgobioworks.com/support/

Digest Run a gel

Ligate Test resistance

From the ‘some PCR

Design primers

Ligate Test resistance

Analyze

<--TRANSFORM

Cloning

Digestions/Ligations

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

Cloning

Gel Electrophoresis ALWAYS USE

LADDERS!!!!

References

Current Protocols in Molecular Biology Search in Pubmed

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

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.