Making BAC Clones and cDNAs Flashcards
What is subcloning?
Moving an insert from one vector to another
What are the types of vectors?
Plasmid vectors
Bacterial artificial chromosomes
What is the purpose of making BAC clones?
Clone all DNA from an organism
How do we make BAC clones?
- Isolate genomic DNA
- Shear DNA into large random pieces
- Ligate into vectors
- Transform
- Plate
What is the result of BAC clones?
A genomic library
Why don’t we use restriction enzymes to make BAC clones?
The DNA fragments would be different lengths
What is the purpose of making cDNAs?
Clone all mRNAs from an organism
What is the procedure to make cDNA?
- Isolate mRNAs
- Convert into cDNAs = synthesis of a complementary DNA molecule
- Ligate into vectors
- Transform
- Plate
What is the result of making cDNAs?
a cDNA library
What vector do we use for human mRNA?
pBluescript because its easiest to work with
How do we find the BAC clone or cDNA we want?
DNA is sequenced before being frozen
What do we use BAC clones for?
FISH
What do we use cDNAs for?
GMOs
