Adept HPLC System 4
High pressure mixing binary gradient system, with UV/Visible detector and PowerStream software.
Adept HPLC system 4
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This binary gradient system uses two Adept CE 4100 high pressure dual piston pumps and the Adept CE 4200 dual wavelength detector for added flexibility.
System control and data processing is provided by 21 CFR part 11 compliance PowerStream data processing software and interface unit, CE 4900. Column heater/chillers can be supplied. The supplied analytical pumps may be substituted by semi-preparative pumps. The CE 4200 UV/Visible detector may be substituted by a Cecil Fluorescence detector. Upgradeable to a ternary gradient system.
Optimisation of a DHPLC Method for Mutation and Polymorphism Detection
This application note by Cecil Instruments describes a method for Denaturing HPLC (DHPLC), an ion-pair reversed-phase high-performance liquid chromatography (IP-RP-HPLC). DHPLC identifies mutations and polymorphisms based on detection of heteroduplex formation between mismatched nucleotides in double-stranded PCR amplified DNA. Sequence variation creates a mixed population of heteroduplexes and homoduplexes during reannealing of wildtype and mutant DNA. When this mixed population is analysed by HPLC under partially denaturing temperatures, the heteroduplexes elute from the column earlier than the homoduplexes because of their reduced melting temperature. Optimisation of DHPLC, requires the generation of a melting profile for each PCR fragment to determine its optimal temperature for analysis.
