Isolation of tetranucleotide loci using 5 - anchored PCR in pineapples (Ananas comosus var. comosus)

Nurafifah Ijap (2005) Isolation of tetranucleotide loci using 5 - anchored PCR in pineapples (Ananas comosus var. comosus). Universiti Malaysia Sabah. (Unpublished)


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This project aims to shed some light on the molecular information of the Ananas comosus var. comosus species, which is a commercial variety of pineapples. This is achieved by isolating tetranucleotide microsatellites, a type of molecular marker which is very useful for a wide range of molecular studies. Therefore, the research objectives were to extract DNA from the pineapple, isolate the micro satellites using 5' - anchored PCR technique with a degenerate primer, PCT6 5'-KKBNVSS(GATA)6-3', submit the isolated micro satellite sequences to the GenBank and obtain Accession numbers for the sequences. The results were five recombinant clones, four successful DNA sequencing reactions, namely PCT 7, PCT 8, PCT 33 and PCT 39, and a total of seven pure tetranucleotide micro satellites isolated from the clones, namely (GATA)n, (CTAT)n, and (TATC)n. Also isolated were one mononucleotide repeat, namely (A)13 and ten cryptic simple regions found in between the 5' and 3' terminal end. The GenBank Accession numbers assigned were AY929614 for locus PCT617, AY929615 for locus PCT618, AY939828 for locus PCT6133 and AY939829 for locus PCT6139 respectively. In conclusion, 5' -anchored PCR technique is highly efficient protocol which saves time and cost, as micro satellites can be isolated without having to perform library enrichment and screening protocols.

Item Type: Academic Exercise
Keyword: Ananas comosus var., pineapple, type of molecular, tetranucleotide, PCR technique, microsatellite
Subjects: Q Science > QH Natural history > QH301-705.5 Biology (General)
Department: SCHOOL > School of Science and Technology
Date Deposited: 31 Dec 2013 11:19
Last Modified: 10 Oct 2017 14:23

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