Amplification and Direct Sequence Analysis of the 23S rRNA Gene from Thermophilic Bacteria. (February 2001)

• Record Type: Journal Article
• Authors: Ibrahim, Ashraf; Hofman-Bang, Jacob; Ahring, Birgitte K.
• Is Part Of: Biotechniques. Volume 30:Number 2(2001)
• Page Start: 414
• Page End: 420
• Publication Date: 2001-02
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/01302rr05
• Languages: English
• ISSNs: 0736-6205

Abstract

We present a simplified and fast method to obtain high-quality sequences directly from PCRs without the traditional gel purification. We also report on an improved method to obtain sequence-quality PCR products from microorganisms that are difficult to lyse with no need for DNA extraction. The technique uses exonuclease I and shrimp alkaline phosphatase to degrade residual dNTPs and primers. Our technique is shown to work on both Gram-positive and Gram-negative bacteria.