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A new 5′ terminal murine GAPDH exon identified using 5′RACE LaNe | SpringerLink

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Molecular Biotechnology

January 2005, Volume 29, Issue 1, pp 39–46 | Cite as

A new 5′ terminal murine GAPDH exon identified using 5′RACE LaNe

  • Daniel Jonathan Park
Research

Abstract

In this work, a ligation-independent, fully gene-specific, nested polymerase chain reaction (PCR) method for the elucidation of 5′ cDNA sequence is described and demonstrated for the first time. Two manifestations of the method, rapid amplification of cDNA ends (RACE) by lariat-dependent nested PCR 5′ (RACE LaNe), at least as simple to perform as conventional RACE, were successfully applied to the murine housekeeping genes phosphoglycerate kinase 1 (PGK1), β-actin (β-ACT), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and the alpha thalassemia mental retardation Y homolog (ATRY) gene of the marsupial, Macropus eugenii. Significantly, a new murine GAPDH 5′ exon, separated by 365 kb of intronic sequence from previously annotated GAPDH sequence, was discovered using 5′RACE LaNe.

Index Entries

5′RACE RACE LaNe GAPDH cDNA sequencing  This is a preview of subscription content, log in to check access.

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© Humana Press Inc 2005

Authors and Affiliations

  1. 1.Department of ZoologyThe University of MelbourneMelbourneAustralia

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