ABSTRACT
Variable amounts of non-specific amplification may occur in HLA PCR-SSP typing, and this can be significantly reduced by the use of AmpliTaq Gold. In an effort to achieve an optimal balance between specificity and efficiency of the PCR amplification for both HLA alleles and the internal control, we designed a system of timed-release activity by combining two different Taq DNA polymerases. The reaction was started at a relatively low level of enzyme activity and as thermal cycling progressed more and more Ampli-Taq Gold was slowly activated for the reaction to continue. We applied this system to all routine HLA PCR-SSP typing. The number of repeat typings due to non-specific amplification and/or amplification failure of the internal control was remarkably reduced.
Subject(s)
Histocompatibility Antigens Class II/genetics , Histocompatibility Antigens Class I/genetics , Histocompatibility Testing , Polymerase Chain Reaction/methods , Taq Polymerase/metabolism , DNA Primers , Histocompatibility Antigens Class I/classification , Histocompatibility Antigens Class II/classification , Reagent Kits, Diagnostic , Time FactorsABSTRACT
PIP: The marriage and family planning counseling duties of the Bereichsarzt (area physician) in the German Democratic Republic are described. He works closely with the marriage and family counseling services of his area as a medical specialist concerned with the physiological and psychological aspects of marriage and family life. He refers patients to the appropriate specialized agencies as a part of his general duties in preventive medicine. In addition, the area physician helps to train counseling personnel in sexual physiology and psychology, equip the counseling center, and coordinate the work of several specialists on a single case.^ieng