Yeast artificial chromosome libraries containing large inserts from mouse and human DNA

Journal Article · Wed May 15 00:00:00 EDT 1991 · Proceedings of the National Academy of Sciences of the United States of America; (United States)

DOI:https://doi.org/10.1073/pnas.88.10.4123· OSTI ID:5934622

Larin, Z; Monaco, A P; Lehrach, H ^[1]

---

1. Imperial Cancer Research Fund, London (England)

Yeast artificial chromosome (YAC) libraries have been difficult to construct with average insert sizes >400 kilobase pairs when DNA is size-fractionated in low-melting-point agarose. By using yeast chromosomes in mock cloning experiments, the authors found that polyamines should be present whenever agarose containing high molecular weight DNA is melted to protect DNA from degradation. By incorporating polyamines during the cloning procedure, they constructed YAC libraries from mouse and human DNA with average insert sizes of 700 and 620 kilobase pairs, respectively. Several genome equivalents of these YAC libraries were replicated onto the surface of many duplicate agar plates using a 40,000 multipin transfer device. High-density filter replicas were screened by hybridization, and 70 mouse YAC clones from 31 loci and 132 human YAC clones from 49 loci were isolated.

• OSTI ID: 5934622
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (United States), Vol. 88:10; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
AMINES
BIOLOGICAL EFFECTS
CHROMOSOMES
DNA HYBRIDIZATION
BIODEGRADATION
DNA SEQUENCING
DNA-CLONING
ELECTROPHORESIS
MAN
MICE
YEASTS
ANIMALS
CHEMICAL REACTIONS
CLONING
DECOMPOSITION
EUMYCOTA
FUNGI
HYBRIDIZATION
MAMMALS
MICROORGANISMS
ORGANIC COMPOUNDS
PLANTS
PRIMATES
RODENTS
STRUCTURAL CHEMICAL ANALYSIS
VERTEBRATES
550200* - Biochemistry