Buenos Dias!  Meeting time for next week?  Undergrad Research Symposium  Reminder – meeting with professors Friday the 16 th from 5-6:30.  Movie Night on Thursday  Real Genius/Caprica – other suggestions?  New Location  Safety Exams!?  Safety Meetings on the 10 th and 17 th  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  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.