Buenos Dias!

1
Buenos Dias!

• Meeting time for next week?
• Undergrad Research Symposium
• Reminder meeting with professors Friday the
  16th from 5-630.
• 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.

2
Lab Protocols
3
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

4
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

5
Cloning

• Biobricks http//ginkgobioworks.com/support/
• Digest
• Run a gel
• Ligate
• Test resistance

• From the some
• PCR
• Design primers
• Ligate
• Test resistance
• Analyze

lt--TRANSFORM?
6
Cloning

• Digestions/Ligations

7
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

8
Cloning

• Gel Electrophoresis
• ALWAYS USE LADDERS!!!!

9
References

• Current Protocols in Molecular Biology
• Search in Pubmed

10
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 its hot)
• List of Supplies

11
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.