Cell and Molecular Biology Lab Final Exam Review Notes

Transformation Experiment

• Marker Genes:

  • Selectable Marker: Beta-lactonase

  • Reporter Marker: Green Fluorescent Protein (GFP)

Plasmids

• Definition: Extrachromosomal circular DNA that replicates independently of the bacterial genome.

• Purpose: Used to amplify and transfer specific genes into organisms (vector).

Phenotype of Successful Transformants

• Characteristics: Colonial growth shows ampicillin resistance and glows green (due to GFP).

• Four Treatments (Plates):

  1. LB-plasmid

  2. LB+plasmid

  3. LB/amp-plasmid

  4. LB/amp+plasmid (look here first for successful phenotype)

Transformation Rate

• Transformation is rare, with a success rate of approximately 1 in 1,000.

Eukaryotic Transformations

• Difficulty: Transforming eukaryotes is more difficult because:

  • DNA must cross both the plasma membrane and enter the nucleus.

  • Bacteria can express plasmid DNA directly, while eukaryotes typically do not.

Critical Steps in Transformation

• Most Critical Step: The 90 seconds heat shock is crucial for transformation success.

Use of CaCl2

• Purpose: Incubate colonies and plasmid in ext{CaCl}_2 to facilitate plasmid DNA proximity to bacterial plasma membrane/wall.

GMO Lab Amplification

• Amplified Genes:

  • CamV Promoter: Derived from Cauliflower Mosaic Virus

  • NOS Terminator: Derived from Agrobacterium Tumefaciens

Meiosis Overview

• Crossing Over: Occurs during Prophase I

• Independent Assortment: Takes place during Metaphase I

• Synapsis Formation: Happens in Prophase I

• Separation of Homologous Chromosomes: Occurs during Anaphase I

• Tetrads Line Up: Happens at the spindle's middle in Metaphase I

• Daughter Cell Chromosomal Makeup:

  • After Meiosis I: Haploid

  • After Meiosis II: Haploid

Cell Cycle

• Stages of Cell Cycle: Understanding what happens in each stage is essential for recognizing cells in mitosis or meiosis based on the number and arrangement of chromosomes.g