Judul: Modification of DNA Extraction from the DNeasy® mericon® Food Kit for Halal Authentication Gelatin
Abstrak: Gelatin adalah bahan yang banyak digunakan di industri pangan, farmasi, dan kosmetik. Status kehalalan gelatin perlu diperhatikan karena dapat bersumber dari hewan babi. Salah satu metode autentikasi halal gelatin adalah real time-polymerase chain reaction (RT-PCR). Analisis RT-PCR memerlukan ekstrak DNA dengan syarat kemurnian A260/280 sebesar 1,8−2,0 dan konsentrasi 10–100 ng μL-1 agar dapat teramplifikasi. Namun, kemurnian dan konsentrasi ekstrak DNA menggunakan standar protokol kit DNeasy® mericon® Food belum memenuhi syarat analisis RT-PCR. Tujuan penelitian ini adalah memodifikasi ekstraksi DNA dari kit DNeasy® mericon® Food. Modifikasi ekstraksi DNA optimal diperoleh dengan kondisi sistem empat kali pooling, unit proteinase K 0,015 AU, waktu inkubasi 180 menit, dan volume larutan EB 30 μL. Kemurnian ekstrak DNA optimal yang dihasilkan pada A260/280 sebesar 1,8 dan konsentrasi 19,4 ng µL-1. Validasi RT-PCR menghasilkan kurva logaritmik dengan nilai Ct 25,5. Hal ini menunjukan ekstrak DNA dapat digunakan untuk autentikasi halal gelatin menggunakan RT-PCR., Gelatin is an ingredient that is mostly used in the food, pharmaceutical and cosmetic industries. The halal status of gelatin needs to be considered because it can be sourced from animals such as pigs. One method of halal gelatin authentication is real time-polymerase chain reaction (RT-PCR). RT-PCR analysis requires a DNA extract with A260/280 purity requirements of 1,8–2,0 and a concentration of 10–100 ng μL-1 in order to be amplified. However, the purity and concentration of the DNA extract using the standard DNeasy® mericon® Food kit protocol did not meet the requirements for RT-PCR analysis. The aim of this research is to modify DNA extraction from the DNeasy® mericon® Food kit. The optimal DNA extraction modification was obtained with four pooling system conditions, proteinase K units of 0,015 AU, incubation time of 180 minutes, and EB solution volume of 30 μL.The optimal DNA extract purity produced in A260/280 was 1,8 and a concentration of 19,4 ng µL-1. RT-PCR validation produced a logarithmic curve with a Ct value of 25,5. This shows that DNA extract can be used to authenticate halal gelatin using RT-PCR.
Keyword: authentication, extraction, gelatin, halal, RT-PCR

Judul: Identification of Pork DNA Fragments in Gelatin Samples Using Real Time Polymerase Chain Reaction (qPCR) methods
Abstrak: Gelatin merupakan suatu jenis protein yang diperoleh melalui hidrolisis parsial kolagen dari kulit dan tulang hewan yang memiliki banyak fungsi untuk keperluan berbagai industri. Kebutuhan gelatin dalam negeri umumnya dipenuhi secara impor. Gelatin yang berasal dari kulit babi adalah yang tertinggi dari total produksi gelatin di dunia. Produk babi dan turunannya termasuk gelatin babi dilarang untuk dikonsumsi oleh Muslim, sehingga diperlukan adanya pengujian untuk mengetahui sumber gelatin. Penelitian ini bertujuan untuk mengidentifikasi fragmen DNA babi pada sampel gelatin dengan menggunakan metode real time Polymerase Chain Reaction (qPCR) dengan primer dan probe sudah termasuk ke dalam master mix mericon Pig Kit. Sebanyak 53 sampel gelatin diekstraksi sesuai dengan prosedur standar dari DNeasy mericon Food Manual Book. Hasil ekstraksi sampel gelatin dikuantifikasi menggunakan nanodrop spectrofotometer 2000c dan diperiksa menggunakan kit komersial. Proses amplifikasi dilakukan sesuai dengan prosedur standar dari DNeasy mericon Pig Kit Handbook. Hasil uji qPCR mampu mengidentifikasi adanya fragmen DNA babi pada sampel gelatin yang ditunjukkan dengan nilai Ct<45. Sebanyak 4 sampel gelatin dari 53 sampel positif mengandung DNA babi, sedangkan 49 sampel gelatin menunjukkan hasil negatif. Uji qPCR dapat dijadikan sebagai metode pengujian untuk mengidentifikasi adanya fragmen DNA babi pada sampel gelatin., Gelatin is a protein obtained by hydrolysis of collagen from animal skin and bone that has great importance in many industry. Domestic gelatin is generally obtained by import. The raw materials used in gelatin production are highly from porcine sources. Porcine gelatin is forbidden for Muslim. For this reason, analytical methods to detect gelatin are needed to make sure the source of gelatin. The aim of this research was to identify the presence of pork DNA fragments in gelatin by real time PCR (qPCR) methods. The primer and probes of qPCR were included in mericon Pig Kit master mix. The samples consisted of 53 gelatins. They were extrated according to the procedure of DNeasy mericon Food Manual Book. The result of extracted gelatins was quantified by nanodrop spectrofotometer 2000c and examined by a commercial kit. The amplification process of gelatins was carried out according to the procedure of DNeasy mericon Pig Kit Handbook. The result showed that gelatins with pork DNA can be identified by Ct value<45. There were 4 samples positive contained pork DNA, while 49 samples were negatives. The qPCR methods can be used to identify the presence of pork DNA fragments in gelatin samples.
Keyword: DNA, gelatin, pork, qPCR

Judul: Thermal Characteristics and Dynamics of Tropical Cyclone over Western North Pacific Ocean: A Case Study of Typhoon Songda (Chedeng) 2011
Abstrak: The frequent occasion of Tropical Cyclone (TC) over Western North Pacific (WNP) Ocean lies from 10˚N to 26˚N and 121˚E to 170˚E. Sources of TC development come from warm ocean temperature where latent heat is released during condensation processes and subsequently concentrated in the boundary layer. Relative humidity at 850-700 hPa is responsible to produce the most intense storm. Meanwhile, weak vertical wind shear between 850-200 hPa is one of the dynamical factors that lead to the development of TC. Pressure level of 850-200 hPa is chosen to minimize the thermodynamic effect in the lower troposphere. Typhoon Songda (Chedeng in the Philippines) or TSC lasted from May 18th to 30th, 2011 and strengthened into powerful Super Typhoon stage between May 24th and 27th. Long curving track and the exhibition of all sequences of TC life cycle, TSC is then considered as an ideal case of TC. The objective of the study with case of TSC is to investigate the role of thermal characteristics and dynamics on TC intensity. The research is carried out by using Advanced Research of Weather Research and Forecasting model (WRF ARW model) version 3.3. Two domains are employed in the model. The first domain situate from 02˚N to 40˚N and 118˚E to 149˚E and cover spatial resolution () of 33.1 km. The second domain situate from 09˚N to 27˚N and 119˚E to 132˚E with of 11 km. Timestep of the model () is 120 seconds. The model is supported by the following physical parameterization, i.e., Yonsei University Scheme (YSU) in the boundary layer, Kain-Fritsch scheme (KF) in the cumulus, and WRF Single-Moment 3-class scheme in the microphysics. Initial meteorological condition prior to the formation of the cyclone is generated using WRF ARW standard-release model. Sensitivity of the model output is obtained from a selected single-parameter change scenarios, namely, 10 % to 20 % decrease and increase of wind shear initial data at 850-200 hPa, and by 10 % to 20 % decrease of relative humidity initial data at 850-700 hPa. The result showed that the negative correlations between wind shear and intensity are found in wind shear scenarios. Wind shears of 10 ms-1 which lead to strengthening of the wind intensity in the early period of Typhoon development are found in wind shear scenarios. The effect of wind shear changes on wind intensity is statistically significant in the 20 % wind shear scenarios. During mature stage, weak wind intensity occurred when wind shear increase as given in the WRF standard-release and the 20 % decreasing scenario. Different situation is found in the 20 % increasing scenario where wind shear tend to weak and lead to strong wind intensity at this stage. Meanwhile, the reduction of moisture supply at 850-700 hPa resulting from 20 % decreasing relative humidity play a major role in the weakening of wind intensity and reduces mechanical energy of the cycle process as much as 300 J kg-1.
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