3.10-3.11 Molecular Biotech Flashcards
Know the basics of gel electrophoresis
DNA has a negative charge so it will migrate toward the + electrode
How is LacZ used in cloning
Bac colonies that contain LacZ will be blue wen grown in media containing X-gal. When lacZ is interrupted by an inserted DNA fragment the colonies will be white
How is LacZ used in cloning
LacZ can be used to screen bac containing recombinant plasmids
What are the basics of PCR
DNA template, DNA pol enzyme, primers, nucleotides, and reaction buffer
What are the weaknesses and strengths of PCR
Weaknesses: Prior knowledge of at least part of the seq of the target DNA. Capacity of PCR to amplify small amounts of DNA, Taq cant proofread. Strengths: Amplify small amounts of DNA from crime scenes. Identify genetic variation. Isolate DNA from ancient sources.
What are the basics of quantitative PCR
probes with dif spectral wavelengths can be used to allow the quantitation of multiple targets in a single reaction
What is chromosome walking and jumping
Walking: sequencing fragments from a point near to a gene of interest. Jumping: large fragment is circularized and the junction is sequenced
What is positional cloning
isolating genes on the basis of their position on a genetic map
How is positional cloning used to clone the CF gene
CF was located by positional cloning
What is a humanized animal model
animal model that carries functional human genes cells tissues and or organs
What is the basic concepts of CRISPR and Cas system of gene editing
CRISPER spacers recognize and cute exogeneous genetic elements by delivering the Cas9 protein and appropriate guide RNAs into a cell the target organisms genome can be cut at any desired location
