Toward a Whole cDNA Catalog

Construction of an Equalized cDNA Library from Mouse Embryos

Naomi Takahashi, Minoru Ko

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

The construction of a cDNA library containing all genes without redundancy is one of the major technical challenges for biology. Toward this goal, we have developed an equalized (normalized) cDNA library from mRNA pools derived from mouse embryos that cover the entire period of mouse ontogenesis. Colony hybridization analyses with 11 genes showed the reduction of abundance variation from at least 6000-fold in the unequalized S-library to ∼33-fold in the EIII-library, which was constructed after three cycles of equalization procedure. Limiting dilution PCR analyses with 26 tissue-specific genes showed the reduction of abundance variation from at least ∼1,000,000-fold in the S-cDNA mixture to ∼100-fold in the EIII-cDNA mixture. Based on these analyses, we estimate that at least 15,000 independent cDNA clones are included with little redundancy in the EIII-cDNA library. This will be a useful resource for mouse biology as well as the mouse genome project.

Original languageEnglish
Pages (from-to)202-210
Number of pages9
JournalGenomics
Volume23
Issue number1
DOIs
Publication statusPublished - 1994 Sep 1
Externally publishedYes

Fingerprint

Gene Library
Embryonic Structures
Complementary DNA
Libraries
Genes
Clone Cells
Genome
Polymerase Chain Reaction
Messenger RNA

ASJC Scopus subject areas

  • Genetics

Cite this

Toward a Whole cDNA Catalog : Construction of an Equalized cDNA Library from Mouse Embryos. / Takahashi, Naomi; Ko, Minoru.

In: Genomics, Vol. 23, No. 1, 01.09.1994, p. 202-210.

Research output: Contribution to journalArticle

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