Methods / Real-time PCR applications
Efficient synthesis of double dye-labeled oligodeoxyribonucleotide probes and their application in a
<%urltitle%>
A fast cleaving non-nucleosidic tetramethylrhodamine dye-labeled support has been developed for automated synthesis of double dye-labeled oligodeoxyribonucleotides in high yield. A mixture (1:1:2) of t-butylamine:methanol:water is used for cleavage and deprotection of dye-labeled oligodeoxyribonucleotides without any degradation or modification of dyes and nucleobases. The cleavage rate of oligodeoxyribonucleotides is significantly increased by using a diglycolate ester linkage instead of the commonly used succinate linkage. These double dye-labeled probes are used in PCR for real time detection of a specific PCR product. Using a 5'-exonuclease assay, detected on the ABI PRISM 7700 Sequence Detection System, there was no distinguishable difference in performance of probes synthesized using the dye-labeled support compared with traditional post-synthetic attachment of rhodamine.
Nucleic Acids Research.1998:26(4):1026
Last update 30-Nov-1999, Rating n/a of 0 votes.
Write your comment
Please Login or Register to Post
Linear-After-The-Exponential (LATE)–PCR: An advanced method of asymmetric PCR and its uses in quan
Rapid and Sensitive Method Using Multiplex Real-Time PCR for Diagnosis of Infections by Influenza A
Comparison of the Real-Time PCR Method and the Gen-Probe Amplified
A novel real-time quantitative PCR method using attached universal template probe
Real-time PCR-based method for the estimation of genome sizes
Real-Time PCR Method for Detection of
Sensitive and Specific Method for Rapid Identification of
Two Novel Real-Time Reverse Transcriptase PCR Assays for Rapid Detection of Bacterial Contamination
Development of a Real-Time PCR Assay for Rapid Detection and Quantification of
Rapid Discrimination between Human Immunodeficiency Virus Type 2 Groups A and B by Real-Time PCR