Designing and testing of primers for cloning of cinnamoyl-coA reductase (CCR) genomic sequence from kelampayan (Neolamarckia Cadamba)

Sia, Jennie. (2010) Designing and testing of primers for cloning of cinnamoyl-coA reductase (CCR) genomic sequence from kelampayan (Neolamarckia Cadamba). [Final Year Project Report] (Unpublished)

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Abstract

Lignin content in the wood is an important criterion to choose planting materials for economic purposes. A study of one of the lignin biosynthesis genes, cinnamoyl-CoA reductase (CCR) from Neolamarckia cadamba was carried out. Partial CCR DNA fragment was amplified by using the primer set 1 designed from partial cDNA of Neolamarckia cadamba. The amplification product was then examined on 1.5% agarose gel and continued the amplified product using primer set 1 did not show homology to the CCR gene from Neolamarckia cadamba. Therefore, primer set 2 was designed from partial cDNA of Neolamarckia cadamba and tested to amplify the target region of the CCR gene. Polymerase Chain Reaction (PCR) optimization of CCR gene from Neolamarckia cadamba was performed by using primer set 2. The PCR product was then purified and cloned into pGEM®-T Easy Vector. However, the estimated product size of the colony PCR of the CCR was not observed positive clone was not obtained. Few solutions could be taken in order to successfully clone the desired PCR product into the pGEM®-T Easy Vector.

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