Growth of Saccharomyces cerevisiae and Preparation of DNA.
Cold Spring Harb Protoc
; 2020(10)2020 10 01.
Article
in En
| MEDLINE
| ID: mdl-33004551
ABSTRACT
This protocol describes methods for isolation of total DNA from a strain of Sacchromyces cerevisiae carrying a recombinant yeast artificial chromosome (YAC). This method is appropriate for preparing DNA that will be subjected to regular agarose gel electrophoresis, Southern blotting, subcloning, genomic library construction, polymerase chain reaction (PCR), or other methods that do not require intact high-molecular-weight DNA. Because the linear YAC DNAs are sensitive to shearing forces, pipettes with wide-bore tips should be used to transfer DNAs. Drop dialysis should be used to exchange buffers. The expected yield from a 10-mL culture is 2-4 µg of yeast DNA.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Saccharomyces cerevisiae
/
DNA, Fungal
/
Blotting, Southern
/
Polymerase Chain Reaction
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Chromosomes, Artificial, Yeast
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Electrophoresis, Agar Gel
Language:
En
Journal:
Cold Spring Harb Protoc
Year:
2020
Document type:
Article