Abstract
This unit describes procedures for generating long-range restriction maps of genomic DNA and for analysis of large insert clones. The basic protocol details restriction digestion of agarose-embedded DNA, PFGE separation, Southern transfer, and hybridization. Support protocols describe the preparation of high-molecular-weight genomic DNA samples in agarose blocks and in agarose microbeads, respectively. Additional support protocols describe the preparation of DNA size standards from l phage and two yeast species, Saccharomyces cerevisiae and Schizosaccharomyces pombe. An alternative method of preparing S. cerevisiae size standards using lithium dodecyl sulfate (LiDS) solubilization is provided. The final protocol details the preparation of BAC DNA suitable for digestion, mapping, and sequencing.
| Original language | English (US) |
|---|---|
| Journal | Current protocols in human genetics / editorial board, Jonathan L. Haines ... [et al.] |
| Volume | Chapter 5 |
| State | Published - Feb 2002 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Medicine(all)
Cite this
Pulsed-field gel electrophoresis for long-range restriction mapping. / Gemmill, Robert M.; Bolin, Richard; Albertsen, Hans; Tomkins, Jeff P.; Wing, Rod A.
In: Current protocols in human genetics / editorial board, Jonathan L. Haines ... [et al.], Vol. Chapter 5, 02.2002.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Pulsed-field gel electrophoresis for long-range restriction mapping.
AU - Gemmill, Robert M.
AU - Bolin, Richard
AU - Albertsen, Hans
AU - Tomkins, Jeff P.
AU - Wing, Rod A
PY - 2002/2
Y1 - 2002/2
N2 - This unit describes procedures for generating long-range restriction maps of genomic DNA and for analysis of large insert clones. The basic protocol details restriction digestion of agarose-embedded DNA, PFGE separation, Southern transfer, and hybridization. Support protocols describe the preparation of high-molecular-weight genomic DNA samples in agarose blocks and in agarose microbeads, respectively. Additional support protocols describe the preparation of DNA size standards from l phage and two yeast species, Saccharomyces cerevisiae and Schizosaccharomyces pombe. An alternative method of preparing S. cerevisiae size standards using lithium dodecyl sulfate (LiDS) solubilization is provided. The final protocol details the preparation of BAC DNA suitable for digestion, mapping, and sequencing.
AB - This unit describes procedures for generating long-range restriction maps of genomic DNA and for analysis of large insert clones. The basic protocol details restriction digestion of agarose-embedded DNA, PFGE separation, Southern transfer, and hybridization. Support protocols describe the preparation of high-molecular-weight genomic DNA samples in agarose blocks and in agarose microbeads, respectively. Additional support protocols describe the preparation of DNA size standards from l phage and two yeast species, Saccharomyces cerevisiae and Schizosaccharomyces pombe. An alternative method of preparing S. cerevisiae size standards using lithium dodecyl sulfate (LiDS) solubilization is provided. The final protocol details the preparation of BAC DNA suitable for digestion, mapping, and sequencing.
UR - http://www.scopus.com/inward/record.url?scp=44049099467&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=44049099467&partnerID=8YFLogxK
M3 - Article
C2 - 18428330
AN - SCOPUS:44049099467
VL - Chapter 5
JO - Current Protocols in Human Genetics
JF - Current Protocols in Human Genetics
SN - 1934-8266
ER -