BACTERIAL TRANSFORMATION AP LAB

Due Tuesday, April 5th or Monday, April 4th by 3:30 for +5 pts!

Write-Up Guidelines

  • Counts as ½ Summative Grade
  • Pre-lab Quiz on Block, March 23rd or 24th = Formative Grade
  • Read Carolina Lab provided AND read Investigation 8 (S97) in your AP lab manual
  • Page 1

 Title

Pre-lab questions – answer questions 1-7 on S-5 & S-6 in Carolina Lab Manual (complete sentences) – Must be completed by Block Day, March 23rd or 24th

  • Page 2

 Abstract

  • Page 3, (4, as needed)

Materials & Methods

  • For a complete listing of materials & methods, see Carolina lab protocol titled, “Transformation”

 Data Analysis

  • Table Must be constructed by Block Day, March 23rd or 24th

Be sure to leave enough room for a detailed description/explanation

  • Purpose & Hypothesis columns for Control Plates must be completed prior to observing plates on Mon, March 28

 Questions

 Error Analysis

…………………………………………………………………………………………

Abstract

  • Single paragraph summary of the lab
  • Should be well-written, informative, and concise
  • ~ 250 words
  • Format
  • Two – three sentences about the topic and its importance
  • Two – three sentences about the actual question you were trying to answer
  • Two – three sentences providing a general description of the procedure used
  • Two – three sentences describing the results
  • Two – three sentences describing your conclusion
  • Which gene(s) were present on the plasmid you used?
  • Justify your conclusion
  • Should provide an excellent explanation of the lab’s principles, procedure, data, and conclusion

Materials & Methods

  • For a complete listing of materials & methods, see Carolina lab protocol titled, “Transformation”

Data Analysis:

  • Plasmid Used: ______

Control Plates

Set-Up / Purpose / “Hypothesis” / # of Colonies Present
LB +
LB –
LB/Amp –
LB/Kan –

Test Plates

Set-Up / Purpose / # of Colonies Present / Appearance of Colonies / Interpretation
LB/Amp +
LB/Kan +

*Please Note: This data table must be constructed by Block Day! Be sure to leave enough room for a detailed description/explanation*

Questions

  • Questions 3-5 must be answered in complete sentences that re-state the question.
  1. Calculate the Transformation Efficiency. Please show your work and record all answers in scientific notation.

 Total Mass of pAMP =

 Total Volume =

 Fraction Spread =

 Mass of pAMP Spread =

 Transformation Efficiency =

  1. If transformation is successful, the antibiotic resistance in the E. coli used in this lab is due to the production of enzymatic protein, which is “built” according to the information encoded in a gene located on the plasmid.

 Sketch and label the basic structure of an amino acid monomer.

 Illustrate a dipeptide formed by showing two amino acids joined together. In addition, provide the following information …

 What type of bond is formed?

 What type of reaction occurs?

 What waste product is formed?

 Specifically, where is this protein construction occurring in the E. coli?

  1. Explain the terms, primary, secondary, tertiary, and quarternary level of protein structure.
  2. Assume an E. coli in your gut is busily synthesizing tryptophan, an essential amino acid for building proteins. You ingest a meal containing a large supply of tryptophan, which is now accessible to the E. coli. Provide a detailed explanation of the bacterium’s response and the mechanism this response represents.
  3. Now assume that same E. coli normally metabolizes the glucose you so generously provide; however, glucose supplies have been mostly depleted. Luckily for your gut E. coli, you drink a large glass of milk. Explain in detail the response that will be seen in the bacterium.

Error Analysis

  • Describe some sources of error in the experimental design
  • Suggestions for improvement