Related Experiment Video
Updated: Feb 7, 2026
Agarose Gel Electrophoresis for the Separation of DNA Fragments
Published on: April 20, 2012
Purification of Yeast Artificial Chromosome DNA for Microinjection Using Pulsed-Field Gel Electrophoresis and
Abstract:
This protocol describes a method for concentrating yeast artificial chromosome (YAC) DNA preparations following separation of chromosomes by pulsed-field gel electrophoresis (PFGE). The agarose slice containing the YAC DNA of interest is melted and digested. The resulting YAC DNA-containing solution is concentrated using ultrafiltration units with a 30,000-mwt cutoff. The DNA is retained in the upper container, where it concentrates sixfold to sevenfold. Ideally, YAC DNA preparations at 10-60 ng/µL can be obtained. However, great care must be taken to recover the YAC DNA molecules that sometimes irreversibly attach to the filter membrane. This procedure can also be used to concentrate BAC DNA from PFGE gel slices of BAC DNA-containing agarose exactly as described for YACs.
Related Concept Videos
02:35DNA Agarose Gel Electrophoresis
Gel extraction follows five major steps: running gel electrophoresis to separate fragments, isolating the individual bands, extracting DNA from those bands, and removing the dye and salts from the extracted mixture to obtain pure DNA.
In cloning experiments, both the insert and vector DNA...
01:22Two-dimensional Gel Electrophoresis
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such...
02:40Chromosome Structure
The centromere is a DNA sequence that links sister chromatids. This is also where kinetochores, protein complexes to which spindle microtubules attach, are constructed after the chromosome is replicated. The kinetochores allow the spindle microtubules to move the chromosomes within the cell during cell division.
Telomeres consist of non-coding repetitive nucleotide...
01:51Lampbrush Chromosomes
LBCs are made up of two pairs of conjugating homologous chromatids. Each chromatid consists of alternatively positioned regions of condensed-inactive chromatin and loosely placed-active side loops, which can be contracted and extended. The loops...
02:31Chromosome Replication
02:04Polytene Chromosomes