Chonnam Med J.  1997 Jun;33(1):131-146.

Optimization of Polymerase Chain Reaction for the Detection of Y Chromosome

• Chung J
• Kim KH
• Oh JS

Abstract

To diagnose X-linked genetic disorders, we optimized the conditions of polymerase chain reaction (PCR) using a pair of primers for the amplification of Y chromosome-specific 198-bps DNA sequence. 1. When cellular DNA was denatured in the reaction, the clear DNA band was detected from 91degreeto 96degreeC irrelevant to the time. 2. The temperature of primer annealing and extension was optimal at 60degreeC but the time didn't affect the result. 3. To improve the reaction, the concentration above 200 M of dNTP each, at 5 and 10pM of primer each and at 1mM of MgCl2 was found to be optimal. The DNA band was detected at the concentration above 1.5 unit of DNA polymerase in 50 l reaction mixture, being distinctive in proportion to the increase of concentration. 4. The DNA band was ambiguous in proportion to the decrease of DNA amount, being detected with minimally 0.1ng DNA. For polymerase chain reaction to determine the prenatal sex, the concentration above 200 M of dNTP each, at 5 and 10pM of primer each, at 1mM of MgCl2, and above 1.5 unit of DNA polymerase in 50 l reaction mixture was found to be optimal.

Keyword

Polymerase chain reaction; Y chromosome

MeSH Terms

Base Sequence
DNA
Magnesium Chloride
Polymerase Chain Reaction*
Y Chromosome*
DNA
Magnesium Chloride