OPTIMASI POLYMERASE CHAIN REACTION (PCR) DARI PROMOTER 35S CaMV UNTUK IDENTIFIKASI TANAMAN TRANSGENIK
At present transgenic plant technology develops quickly and it is expected that the distribution in markets will increase. Therefore, it is necessary to develope a method for detecting the genetically engineered products, which can be distinguished from non-engineered ones. The detection can be done...
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| Main Authors: | , |
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| Format: | Theses and Dissertations NonPeerReviewed |
| Published: |
[Yogyakarta] : Universitas Gadjah Mada
2011
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| Subjects: | |
| Online Access: | https://repository.ugm.ac.id/88657/ http://etd.ugm.ac.id/index.php?mod=penelitian_detail&sub=PenelitianDetail&act=view&typ=html&buku_id=50672 |
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| Institution: | Universitas Gadjah Mada |
| Summary: | At present transgenic plant technology develops quickly and it is expected
that the distribution in markets will increase. Therefore, it is necessary to
develope a method for detecting the genetically engineered products, which can
be distinguished from non-engineered ones. The detection can be done through
the identification of 35S CaMV promoter by using the Polymerase Chain
Reaction (PCR) method due to the fact that most transgenic plants currently used
the kind of promoter. Thus, it is necessary to optimize the 35S CaMV promoter
amplification for better results. In this study, the optimation efforts was done for
annealing temperatures and the number of template DNA. The study is started
by the extraction and purification of soybean DNA samples by using the kit
method (PureLinkTM Plant Total DNA Purification Kit) and the manual method
(extraction buffer), followed by the amplification process using a variety of
annealing temperatures and the number of template DNA.
Results of the extraction and purification of the soybean DNA samples in
the study showed that using the kit method, DNA obtained were 0.06% (58.5 μg
from 100.0 mg soybean samples) with a purity level of 1.77, and using the
manual method were 0.04% (361 μg of 1.0 g soybean samples) with a purity level
of 1.88. Result of the optimation of primary annealing temperatures for the
CaMV 35S promoter amplification was 550C, while for the number of template
DNA was 300 ng. Optimum condition of the 35S CaMV promoter amplification
could be used to specifically identify the transgenic soybeans using the samples
of DNAs resulted from either kit method and the manual method |
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