METODE EKSTRAKSI DNA CABAI (<em>Capsicum annuum</em> L.) MENGGUNAKAN MODIFIKASI BUFFER CTAB (CETHYL TRIMETHYL AMMONIUM BROMIDE) TANPA NITROGEN CAIR

Abstract

A B S T R A C TChili pepper is an agricultural commodity having high economic value. The production and supply of chili pepper frequently did not match the increased demand; it caused the market price fluctuated. It is important to create new varieties of chili pepper with high production trait to overcome the scarcity. Therefore the plant breeding activities for chili pepper should be done intensively in both conventional and molecular-based to obtain varieties of chili pepper with expected qualities. In molecular breeding, DNA extraction is the crucial steps of the process. If extracted DNA has an excellent quality and quantity, the next processes normally could be completed with the high-quality result. To date, most methods of DNA extraction used liquid nitrogen to destroy the tough carbohydrates of plant tissue. Liquid nitrogen is nitrogen gas in a fluid state which quite difficult to be distributed to the remote laboratory wit no available storage facility. This study aimed to obtain a modified DNA extraction method, in particular for chili pepper, which capable to produce DNA with high quality and quantity without using liquid nitrogen. The sample used consisted of eight F2 plants including their hybrid-parental of the Kencana and the 0207. This research applied modified Doyle and Doyle method for extraction. Modification of extraction buffer is done through the addition of the 1% (w/v) PVP (Polyvinylpyrrolidone) and 0.2% (v/v) β-mercaptoethanol. The results showed that the DNA extracted using this method has good quality and quantity, capable of being amplified by using SSR (Simple Sequence Repeat) primer and could be digested by restriction enzyme EcoRI. Besides, this method can reduce dependence on the use of liquid nitrogen, in particular for remote laboratories with no available storage facility.

Multiplex PCR Method of Detecting Pork to Guarantee Halal Status in Meat Processed Products

“…This was due to the extracted samples coming from different sources there was meat and meatballs and sausage. According to the KapaBiosystem (2014) recommendation cited by Nugroho et al (2017), the template DNA concentration needed for PCR activities ranged from 10-100 µg/mL whereas according to Maryam (2014) the optimal concentration in PCR amplification at 30 cycles to produce thick bands was a 50 µg/mL concentration both in beef comparison, pork, beef meatball samples and pork meatball samples. In this study showed that the concentration of DNA in beef and pork samples was very high, namely 558.90 and 610.65 µg/mL, because the extracted sample was a fresh meat without any mixture material.…”

Validation of molecular markers associated with amylose content of F8 and F9 lines resulting from a crossing of Black Rice and Mentik Wangi

“…DNA extraction was carried out using the Cetyl Trimethyl Ammonium Bromide (CTAB) method [19], which was modified by adding 2% PVP (Polyvinylpyrrolidone) and β-mercaptoethanol. The addition of PVP is carried out to reduce polyphenolic compounds that can bind to DNA, while the compound βmercaptoethanol was added to the extraction buffer to degrade proteins in the sample so that pure and high DNA results will be obtained [20].…”

“…The homogenized DNA solution and DNA Ladder were injected into the well of a 1% agarose gel (0.5 grams of agarose, 50 ml of 1x TBE buffer, and 5 µl of fluorosafe), which was submerged in 1x TBE buffer solution and ran at 50 volts for one hour. After the electrophoresis process was complete, the agarose gel was transilluminated under a UV transilluminator [20]. The purity of the DNA solution was determined if there was a single genomic DNA band and there was no smear on the agarose gel resulting from electrophoresis [17].…”

Design and Performance Test of Specific Primers to Detect Bovine DNA Fragments using Multiplex PCR Technique for Halal Authentification

“…Therefore, DNA isolates from meat samples were tested quantitatively, as shown in Table 2. The concentration of DNA templates required for the PCR process ranges from 10 to 100 ng/μl (Nugroho et al, 2017). It is the total concentration of DNA in the PCR mix.…”