Class notes
BIOB12 Lecture Notes (1-12)
- Institution
- University Of Toronto (U Of T )
BIOB12 Lecture Notes (1-12)
[Show more]
Connected book
Book Title:
Author(s):
- Edition:
- ISBN:
- Edition:
Content preview
LECTURE 02 - SyllabusLECTURE 02 - Spectrometer
● A = ecl
○ Absorbance
○ Extinction coefficient
○ Length of light
● Graphing - absorbance vs concentration
○ Slope = extinction coefficient
● Pipettes
○ Blow out/serological - two circles at top, goes to 9mL
○ To deliver/Mohr - no ring at top, goes to 10 mL
○ Volumetric/transfer pipette - exact amount
● DNA - OD260
● Protein - OD280
● Growth curve - lag phase, log phase, stationary phase, death phase
○ High turbidity = high absorbance = low transmittance = many cells = either dead or alive
LECTURE 03 - cell growth
● Bacteriostatic - does not kill organisms, inhibits reproduction
● Bactericidal - kills organisms by inhibiting cell wall synthesis
● Colony counting
○ Less than 20 = TFTC
○ More than 300 = TMTC
LECTURE 04 - molecular cloning
● 3 distinct properties of a vector
○ Origin of replication - able to replicate independently of host
○ Restriction enzyme site - allow target foreign DNA to enter
○ Selectable marker gene - select for cells that can take up vector
● Competent - cell capable of taking up vector
● Transformant - host that has taken up vector
● Recombinant vector - foreign DNA inserted
LECTURE 05 - DNA sequences
● 5’ overhang has a short 5’ end
● 3’ overhang has a short 3’ end
● Blunt ends has equal length cut fragments
LECTURE 06 - Restriction Maps
● Anchor sites - places the enzyme cuts the DNA into fragments, sites
○ Second anchor site - used for another enzyme or to find double digests
● DNA fragments
○ Large - nicked
○ Medium - linear
○ Smallest - supercoiled
● PROCEDURE - RESTRICTION DIGEST
○ Each digest contains
■ Plasmid (recipient DNA holder)
■ Restriction buffer (sink in gel and colour)
■ Sterile water (volume)
■ Restriction enzymes
● EcoRI
● HindIII
● KpnI
● BamHI
○ Tap to mix / briefly spin
○ Incubate in 37C-45C for 35min-45min
○ Already have Anza Red Dye, add bromophenol blue dye if necessary
■ Loading dye and loading buffer will cause restriction enzymes to cut incorrectly
○ Leave for next day
The benefits of buying summaries with Stuvia:
Guaranteed quality through customer reviews
Stuvia customers have reviewed more than 700,000 summaries. This how you know that you are buying the best documents.
Quick and easy check-out
You can quickly pay through credit card or Stuvia-credit for the summaries. There is no membership needed.
Focus on what matters
Your fellow students write the study notes themselves, which is why the documents are always reliable and up-to-date. This ensures you quickly get to the core!
Frequently asked questions
What do I get when I buy this document?
You get a PDF, available immediately after your purchase. The purchased document is accessible anytime, anywhere and indefinitely through your profile.
Satisfaction guarantee: how does it work?
Our satisfaction guarantee ensures that you always find a study document that suits you well. You fill out a form, and our customer service team takes care of the rest.
Who am I buying these notes from?
Stuvia is a marketplace, so you are not buying this document from us, but from seller StudentUofT. Stuvia facilitates payment to the seller.
Will I be stuck with a subscription?
No, you only buy these notes for CA$6.66. You're not tied to anything after your purchase.
Can Stuvia be trusted?
4.6 stars on Google & Trustpilot (+1000 reviews)
50064 documents were sold in the last 30 days
Founded in 2010, the go-to place to buy study notes for 14 years now