酵母人工染色体

 

 

・         Easy YAC Preparation Method (Andrew Davies,Shaw lab)

・         Screening YAC libraries (Donis Keller Lab)
This is a method for screening YAC libraries for specific DNA sequences. Transformants are patched onto selective medium plates and colonies lifted to nylon filters for further growth. Cells are then lysed in place and the DNA is fixed to the filters. DNA probes are hybridized to the filters to test for the presence of specific sequences. A secondary screen is performed to ensure a positive hybridization.  

・         YAC library screening by PCR on three-dimensional DNA pools (Stanford)
Provides procedure for preparation of DNA Pools and PCR Screening

・         YAC library screening by hybridization (Stanford)
Provides procedure for yeast DNA preparation in MT plates, preparation of hybridization filters, hybridization    

• Preparation of YAC End Probes (Stanford)
  Procedure for arm rescue
    
• Gel-Purified, Agarased YACs for Yeast Transformation (Andreas Gnirke)
  A protocol for purifying yeast artificial chromosomes by gel electrophoresis and retrieving the resultant products. The products are suitable for use in transformation experiments
     

・         Full Scale YAC PLUG Preparation (WUGSC)
    

・         YAC Plug 25ml QC Preparation (WUGSC)
    

• Restriction Digests of High Molecular Weight Yeast DNA (Donis Keller Lab)
  Restriction digests of YACs for mapping using rare cutting enzymes or more conventional restriction endonuclease digestion
     
• Restriction Digests of YACs in Agarose Plugs (Donis Keller Lab)
  Restriction enzyme digests on intact, high molecular weight YAC DNA, for use in pulsed-field mapping experiments.
    
• Sizing of YAC Clones (Donis Keller Lab)
  To determine the size of a yeast artificial chromosome within the background of the normal yeast chromosomal complement
    
• Subcloning of Yeast Artificial Chromosomes into Phage Lambda (Donis Keller Lab)
  To subclone the large insert fragment of human DNA contained within a YAC into a bacteriophage lambda vector. 
     
• Identification of End Clones in YAC Subclone Libraries (Donis Keller Lab)
  To identify the YAC subclones containing both a human insert and a portion of either the left or right arm of the pYAC4 vector.