Lecture 38 - Moleculer Genetic Analysis and Biotechnology II Flashcards
How are DNA libraries screened?
L38 S5
- bacterial colonies containing cDNA are grown
- cells from colonies are transferred to filter
- cells are disrupted, exposing DNA
- labeled probes are applied to see if they bind desired sequence
- filter is used to identify bacteria with desired sequence on original plate
What is Sanger’s method of DNA sequencing?
L38 S13-14
Using a low concentration of dideoxyribonucleotides which randomly will terminate replication.
This is done separately with all 4 nucleotide.
Segments can then be separated by size and each fragment length will show which nucleotide is at that position.
What is the Ata skeleton?
L38 S28
A human skeleton with several mutations resulting in a grossly abnormal development
What are humanized mice?
L38 S34
Mouse with functioning human genes making it a better model for study.
Very good for immunologic studies.
What is siRNA and how can it be useful in treating disease?
L38 S39
Small, double stranded DNA
It interferes with mRNA that it is complementary with
Can be used to target certain genes that are overactive or producing a defective product
What is shRNA?
L38 S41
Artificial RNA molecule with a hairpin loop that is similar to siRNA but more stable in the cell
How does CRISPR-Cas9 work?
L39 S3
Uses RNA as a guide for a nuclease which cuts a certain DNA sequence.
This allows for inactivation of that DNA sequence or insertion of a new sequence
