Bio 1AL Lect 2

Bio 1AL Lect 2 - Bio 1A Lab Lecture Outline # 2 Vibrio...

Info iconThis preview shows pages 1–3. Sign up to view the full content.

View Full Document Right Arrow Icon
Bio 1A Lab Lecture Outline # 2 Vibrio streaking & Enzymes February 1 st , 2010 I. Vibrio isolation. A. Plate sea water onto SWC plate (Lab 1) B. Pick bioluminescent colony and streak onto SWC plate (Lab 2) C. Pick bioluminescent colony and streak onto SWC plate (Lab 3) D. Pick bioluminescent colony and set up a PCR reaction (Lab 4) E. Analyze PCR reaction – clean up and run a gel (Lab 5) i) If successful set up sequencing reactions (forward and reverse) F. Data anlysis – consensus sequence, Blast, Clustal (Lab 6) II. Light Spectroscopy A. Overview B. Identification – Absorption Spectrum C. Quantification – (A = clZ) D. Blank – why? III. Protein Structure: A. Amino Acids: classified by R group side chains a. Hydrophobic b. Hydrophilic c. Ionizable: NH2, COOH B. Polymerization C. Levels of structural organization: a. 1 o - primary sequence b. 2 o – hydrogen bonds from backbone molecules c. 3 o - prosthetic groups in addition d. 4 o – requires > 1 subunit IV. Reactions: Substrate(s) Product(s). A. Progress of Reaction: Δ G o , Δ S ± , E a . B. Boltzman – Energy Distribution. a. Heating does not change Δ G o nor E a , but the energy distribution changes. V. Enzymes A. Enzymes speed up chemical reactions by decreasing E a . B. How do enzymes work? a. Local environment, pH, proper orientation, distortion, change strength of binding. C. Isoenzymes (Isozymes). VI Measuring Kinetics A. Substrate Product. B. Direct/Indirect Measurements.
Background image of page 1

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
VII. SalivaryAmylase A. Starch (Amylose) ------------> Maltose. B. Structure of Starch – (polymer α 1,4 glucose), focus terminal glucose: cyclical linear. C. Conversion – what happens: [maltose] & [starch/amylose]. a. Measure maltose indirectly by number of reducing units. D. Measure – Reduction a. Maltose + DNS (yellow) -------> Soluble brown substance; measure O.D. 510 nm b. Glucose – linear (L) vs. ring (R) form. c. amylose – terminal glucose (L) to (R) A (R) --- B (R) --- C (R) -- D (R) --- E (R) ---F (R) A (R) - B (R) - C (R) - D (R) - E (R) -F (L) d. Example: Amylose (chain of 6 Glucose) 3 Maltose 1 reducing unit is converted into 3 reducing units Actual difference = 2 reducing units! We need a blank. VIII. Measuring Kinetics: Part I A. Δ [E] 50ng 400ng a. Excess Substrate. 1 ml of S b. Keep all parameters constant except one variable. B. Experimental Setup: Table on P. 60 a. Blank: 0.5 ml 100ng/ml amylase, 1 ml DNS, 1 ml S (mix stock prior to removing) b. Reaction tubes!
Background image of page 2
Image of page 3
This is the end of the preview. Sign up to access the rest of the document.

This note was uploaded on 02/27/2010 for the course BIO 1al taught by Professor Pederson during the Spring '08 term at Berkeley.

Page1 / 13

Bio 1AL Lect 2 - Bio 1A Lab Lecture Outline # 2 Vibrio...

This preview shows document pages 1 - 3. Sign up to view the full document.

View Full Document Right Arrow Icon
Ask a homework question - tutors are online