Optimisation of polymerase chain reaction (PCR) conditions for the amplification of Rubisco gene (RBCL) in Triticum Aestivum (wheat)
- Author
- Manhimanzi, Washington
- Title
- Optimisation of polymerase chain reaction (PCR) conditions for the amplification of Rubisco gene (RBCL) in Triticum Aestivum (wheat)
- Abstract
- The aim of the study was to optimise PCR conditions for the amplification of the RubisCO gene rbcL from Triticum aestivum (wheat). The DNA extracted from the four Triticum aestivum samples was of very high integrity, this can be noted by sharp distinct bands seen when the samples were run on a 1% Agarose gel. Absence of smears in any of the lanes indicated that no sample was degraded. Unfortunately, the concentration and purity of DNA could not be determined as the Nanodrop Spectrophotometer was down. Hence, DNA concentrations were not optimised for the PCR reaction. The primers bind to the template DNA indicating the specific points at which the polymerase enzyme can start complementary strand synthesis. Higher concentration of primers will result in primer dimers which might affect downstream applications like sequencing. There will be need to clean up the amplicon thoroughly with exonuclease enzyme to ensure sequencing reaction is successful.
- Date
- November 2020
- Publisher
- BUSE
- Keywords
- Polymerase Chain Reaction Conditions
- Amplification of Rubisco gene
- Triticum Aestivum
- Supervisor
- Ms. Muyambo
