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Buenos Dias!

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??

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Buenos Dias!

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  1. 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.

  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 <--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 it’s 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.

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