Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Search research articles

Related Experiment Videos

cDNA selection with YACs

¹Skin Biology Research Center, Johnson and Johnson, Skillman, NJ 08588-9418, USA.

Molecular Biotechnology

|January 23, 1998

Summary

This study presents a novel polymerase chain reaction (PCR)-based method for identifying coding sequences in large genomic DNA fragments. The efficient technique aids in positional cloning and genomic mapping by selecting expressed sequence tags (ESTs).

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Bioengineering the hair follicle.

Organogenesis·2009

Same author

Phylogeny of the hair follicle: the sebogenic hypothesis.

The Journal of investigative dermatology·2007

Same author

What is the biological basis of pattern formation of skin lesions?

Experimental dermatology·2006

Same author

Scd3--a novel gene of the stearoyl-CoA desaturase family with restricted expression in skin.

Genomics·2001

Same author

Asebia-2J (Scd1(ab2J)): a new allele and a model for scarring alopecia.

The American journal of pathology·2000

Same author

Marie Unna hereditary hypotrichosis gene maps to human chromosome 8p21 near hairless.

The Journal of investigative dermatology·2000

Area of Science:

Genomics
Molecular Biology
Biotechnology

Background:

Identifying coding sequences within large genomic segments is crucial for positional cloning and genomic mapping.
Existing methods may lack efficiency or simplicity when dealing with extensive DNA fragments.

Purpose of the Study:

To develop a simple and efficient polymerase chain reaction (PCR)-based approach for identifying coding sequences in large genomic DNA fragments.
To facilitate the identification of expressed sequence tags (ESTs) for genetic studies.

Main Methods:

A PCR-based strategy was employed using genomic DNA cloned in yeast artificial chromosome (YAC) vectors.
Repetitive and GC-rich sequences were blocked in immobilized genomic DNA.
Hybridization with cDNA libraries followed by PCR was used to recover selected coding sequences.

Main Results:

The method successfully identified coding sequences in large genomic fragments.
The procedure demonstrated efficiency and simplicity.
The technique was validated using single or multiple cDNA libraries and total yeast DNA containing a YAC.

Conclusions:

The described PCR-based approach provides an efficient means to identify expressed sequence tags (ESTs) in large genomic DNA.
This method is valuable for positional cloning and genomic mapping studies.
The technique is adaptable and effective even with complex DNA constructs like YACs.

Related Experiment Videos