Establishment of experimental conditions on Random Amplified Polymorphic DNA (RAPD) analysis of Lansium domesticum Corr. I. DNA extraction from leaf samples
การศึกษาสภาพที่เหมาะสมในการใช้เทคนิคอาร์เอพีดีสำหรับพืชสกุลลางสาด (Lansium
domesticum Corr.) I. การสกัดดีเอ็นเอจากตัวอย่างใบ
Charassri Nualsri, Asst. Prof., D. of Plant Sci., F. of Natural Resources, PSU.
Suvimon Konlasuk, Student
Grant : Prince of Songkla University
Published : Songklanakarin J Sci Technol 2000, 22(4) : 403-410
Key words : RAPD, PCR, Lansium domesticum, DNA extraction
DNA extraction from leaf samples of longkong (Lansium domesticum Corr.) was investigated. From four different methods, the CTAB method modified from Doyle and Doyle (1990) showed the best result. From 200 mg fresh leaves as starting material, approximately 15-20 mg of DNA was obtained. DNA extracted from leaves in three different stages of development was compared. Young leaves tended to produce higher yields while young fully expanded leaves and old leaves gave almost the same amounts of DNA. Quantity and quality of extracted DNA from leaves that were kept in a
sealed brown paper bag and stored in various durations under 3 different conditions, room tempera-
ture (30-32oC), 4oC and -30oC, were determined. At room temperature and 4oC, DNA yields declined rapidly after storage for 3 and 5 days, respectively, while leaves storage under -30 oC could be kept at least 30 days and the banding profiles of RAPD-PCR products was not different from fresh leaf extracted DNA.
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