Who knows we could learn from unconquerable and dangerous open ocean. the deathful thing with huge wave that will smash and role you up and down very harshly. this certainly drives you into paralyzing thought since you can’t handle that rapid motion with your hand and leg. There is no way to escape from that threat once you swim on the open ocean surface. To what’s more, there will be also frightening threat may come from blue-blood killer creatures with their unpredicted and rapid movements that my come from behind, underneath or in front of you while swimming. They might be appearing and eating you alive with no sign. Once you catch their attention it will preys you. At that point you are in emergency as if no life tomorrow.

This perception is never true for Bakhti Sarma, athlete from Rajasthan, who set record on long-distance swimming in Antarctic ocean. In one-to-zero degree water she could swim and spent quite long time there easily. To her, ocean is the play arena by means it brings her to joy and passion.  Because of that, swimming is the enjoyable activity which she could spend many hours. Based on her explanation, to acquire the skill she had to swim 9 hours in average everyday beside doing chores, study, and other social activities like normal people. Only she sacrificed a lot more time for swimming despite playing with friend representing her consistency and perseverance. Though sometimes she comes into think that she needs quit and give up on training because of the calmness and loneliness she witnesses. Accordingly, for the raise of her life she had spent 15 years swimming. For that long time period she has experienced no friend as no any chance to make friend. The better companion she gets is herself with thought might crossing. The things that may not be seen but support her every time she face hard situation while swimming. More importantly, it changed her personality into resilience toward dangerous threatening may come every time facing open ocean.

It’s countless for her facing difficulties especially when it comes to break the records. Only those who are well-trained with myriad exercises could win the award. For example, when she had to win award to swim in arctic ocean. she conquered the dense and painful freezing ocean while nobody would do that insane record. With her sense of completion coming from the voice within that kept her on fire for every moment, she stroked and kept her body moving forward diminishing demotivated feeling or the thought to give up. To ignore that, she kept thinking that her struggling need to be paid off despite negative thought that haunted her mind. Every time she felt physical paralyzing, once said you don’t good enough to win this record and you are not well-trained to beat this. Yet, she knew her capability and trust the positivity coming from inside. In that difficult situation it could be able to make a joke. One time, while doing some strokes there was penguin swum underneath and moved to her side the think that crossed her mind was “The penguin cheered you up”. They swam together for 15 minutes and set the unbelievable record. Surprisingly, there is even no any snow in Rajashtan since it’s area on earth. Yet her determination and confidence leads her to set long-distance swimming in extreme Antarctic temperature.

What we can learn from this true story is the only limit you have is yourself. There is nothing that could lay yourself down other that you. Who knows the girl from arid area Rajashtan could swim for long time period in Antarctic ocean. all she has is that stone hard intension and from deep inside yourself together with persistency on training and exercising.

One of critical global issue we’re facing today is insufficient of substantial crop production. it contains many necessary matter to support our body mechanism including carbohydrate, protein and triglyceride. These particular maters are so essential and store massive energy resource to generate live for every individual. Lack of these would drive to body abnormalities especially slow rate metabolism and eventually leading to organ dysfunctionality. many people in developing countries has witnessed this such of disease. All these things happened because of low food production.

Despite of its insufficiency, based on data reported by FAO, the annual disposal number of food is reaching at roughly 6 million tons. This might be coming from restaurant disposal or from household that being spoiled or expired on fridge on the day-to-day basis. We could found these on the way nearby restaurant or urban wasteland and very congestive to some area of the city. To address this issue, we could exemplify on dump area in the suburb or inner-city are which is putrefied because of this spoiled food. thus, not only to improve the agriculture productivity there should also the need of food management to diminish of its disposal.

It has been projected that the demand of food to feed 6 billion populations would be increased by three times for over 3 decades ahead. Unless the intention to fulfill this number there will never be enough for global food to feed people. Traditional farming alone would exactly be suppressed to produce sufficient food. This situation might lead to collapse because imbalance between food and number of people occupy on earth. For some reason, we should aware to this projection especially people who work in agricultural sector.

There are many ways to improve production more achievable is intensification. There is long-lasting research that putting together massive experiment to fulfill the demand on food globally. Currently, researchers are now attracted to investigate more about so called “climate or weather-proof farming”. As it is named, this type of farming is rather not affected by outside-world factors such as wind, temperature, humidity and light intensity etc. it is very identical to growing plant on vertical rack together with controlled environmental system. When it comes to the growing medium, instead of using soil, they used artificial substitute such as polyurethane sponginess or biodegradable peat moss. Under sophisticated control by highly smart robotic system the nutrient formula as well as the environment is control thoroughly by fascinating technology. With this tools the nutrient is constantly and consistently pumped through this facility and targeted directly into plant root zone.

This technology is insane. Instead of using artificial light the expert completely supports this farming method with fiber-optic cable. this innovation channels photon from daylight into the receptor located inside the plantation area to promote plant growth. Moreover, unlike traditional viticulture, this farming could be applied in unpredictable place such as on decommissioned container ship, lobby hotel or underutilized room in house. More importantly, there is no any obstacle in growing plant. That being said, we could cultivate the plant anywhere any time. Under no circumstance could we grow the vegetables or herbs in unfavorable condition since it’s automatically controlled and configured to support plant growing. Struard oda, An Indoor vertical farming practiciant, said that by applying this technique would get year-round vegetable production with consistent quality and predictable output. Putting together this is the effective way to supply food for family or even regionally.

Henceforth, there could not be an obstacle in providing food as this technology fully optimized and provided food for family. This is the farming revolution method answers the need of sufficient yet good quality food.

ISOLASI DNA DAN PURIFIKASI

Penelitian dasar dalam bioteknologi adalah kemampuan memanipulasi DNA. Sebelum pengerjaan DNA rekombinan, peneliti membutuhkan sebuah metode dalam mengisolasi DNA dari organisme. Mengisolasi DNA dari bakteri sangat mudah karena sel memiliki struktur dinding sel dan membran sell. Bakteri seperi E. Coli merupakan organisme yang lebih diutamakan terkait manipulasi genetic karena mudah mengisolasi DNA nya. E. coli menjaga DNA genomic dan plasmidnya dalam. Sedangkan ukuran keduanya berbeda sehingga mudah dipisahkan.

Untuk membebaskan DNA dalam sel, sel membrane harus dihancurkan terlebih dahulu. Apda bakteri, enzim Lysozyme menghancurkan peptidoglikan yang menyusun dinding sel. Kemudian, detergen menghacurkan membrane sel dengan memecah dua lapisan lipid. Untuk organisme lain berbeda tergantung senyawa penyusunnya. Sel hewan dan tanaman harus terlebih dahulu digiling untuk membebaskan komponen dalam sel. Sel tanaman harus dihancurkan secara mekanik untuk merusak dinding sel, kemudian jaringan dinding dipecahkan dengan enzim untuk memutuskan ikatan polimer menjadi monomer.

Ketika komponen intra seluler dipisahkan dari struktur luar dengan sentrifugasi atau dengan ekstraksi kimia. Centrifugasi dapat memisahkan komponen berdasarkan ukuran, karena laju pengendapan molecule besar akan lebih cepat disbanding molekul kecil. Contohnya, setelah dinding sel di pecahkan, ukuran fragmennya akan bervariasi. Sentrifugasi akan menyebabkan DNA membentuk pellet, numun dinding sel yang terlarut akan tetap dalam larutan. Ektraksi kimia menggunakan phenol untuk menghilangkan protein yang tidak diinginkan seperti Histon. Pheol adlaah asam yang larut dalam 60 – 70% senyawa organic terutama protein. Phenol tidak larut dalam air, dan ketika dicampurkan dengan air sample DNA dan protein, akan menyebabkan dua fraksi yaitu air dan seperti minyak. Protein akan larut dalam lapisan phenol dan DNA pada lapisan cair. Dua fasa dipisahkan dengan sentrifugasi dan lapisan DNA dipindahkan dari phenol. Ketika protein dibuang, sampel masing mengandung RNA besama DNA. Dengan menggunakan enzim RNAase RNA akan degrasasi.

Eletrophoresis Memisahkan fragmen DNA berdasarkan ukurannya

Elektroporesis Gel beserta pewarnaan dengan ethidium bromide digunakan untuk memisahkan fragmen DNA berdasarkan ukurannya. Gel pada elektroporesis Gel mengandung agarose, yaitu ekstrak polisakarida dari rumput laut yang bersifat seperti gelatin. Agarose merupakan bubuk yang larut dalam air hanya ketika dipanaskan. Seletelah dingin, agar akan mengeras. Untuk visualisasi DAN, agarose dibentuk pada cetakan persegi yang disisipkan sisir pada salah satu ujung agar untuk membuat sumur atau lobang. Setelah gel memadat, sisir akan diangkat dan membentuk sumur.

Elektorforesis gel menggunakan listrik untuk memisahkan molecule DNA berdasakan ukurannya. Agarose direndam pada tangka yang memiliki eletroda positif pada satu ujungnya dan negative pada ujung lainya.

DNA sample diloading kedalam sumur/ lobang kemudian dialirkan arus listrik. DNA akan bermigrasi melalui gel. Fosfat DNA bermuatan negated jadi akan berpindah dari negative ke positif, agarose yang dipolimerisasi bertindak seperti saringan dengan lobang yang sangat kecil antara rantai polimer agar. DNA bermigras melalui pori ini. Agarose memisahkan DNA dengan berdasarkan ukuran karena bagian yang besar akan bermigrasi lebih lambat.

Untuk memvisualisasikan DNA, gel agarose akan dihilangkan dari tank kemudian direndam kedalam larutan etidium bromide. Pewarna ini akan membentuk interkalasi antara basa DNA/ RNA. Molekul DNA pada ukuran yang sama pada umumnya dari pita yang tebal dan ukurannya akan dapat ditentukan dengan membandingkan dengan standar berat molekul yang diloading pada wel yang berbeda. Karena standar tersebut diketahui ukurannya secara spesifik.

Ukuran DNA yang diuji mempengaruhi jenis gel yang digunakan. Standarnya adalah agarose, namun DNA yang ukuran kecil dari 50 hingga 1000 bp lebih baik digunakan polyacrylamide. Gel ini dapat memisahkan fragmen DNA yang berbagai macam. Untuk ukuran yang sangat besar (10.000 – 10 jt bp) agarose digunakan namun arus digantikan dnegan dua arah. Pulsed Field Gel Electrophoresis (PFGE). Setiap perubahan arah meloloskan DNA ukuran besar yang tersangkut dalam pori agar. Kemudian menyebabkan meraka bermigrasi lebih jauh. Akhirnya elektroforesis dapat memisahkan fragmen yang memiliki selisih yang kecil.

Enzim Restriksi Memotong DNA, Ligasi menyambungkan DNA

Kemampuan untuk mengisolasi, memisahkan dan memvisualisasi fragmen DNA tidak akan berguna jika tidak ada metode untuk memotong DNA kedalam ukuran yang bervariasi. Pada kenyataanya, Kejadian ini terjadi secara natural dengan bantuan enzim restriksi atau restriction endonucleases merupakan kuncinya. Enzim bakteri ini dapat berikatan dengan sisi pengenalan spesifik pada DNA dan memotong pada kedua untai DNA. Ini berevolusi untuk melindungi bakteri dari DNA asing, seperti Virus. Enzim tidak memotong DNA nya sendiri karena mereka dimetilasi, yaitu jika Sekues pengenalan dimetilasi, maka enzim tidak dapat berikatan.

Perbedaan pemotongan menentukan jenis enzim restriksi. Enzim restriksi tipe I memotong DNA 1000 atau lebih pasang basa dari sisi pengenalannya. Tipe II memotong ditengah-tengah sekuens pengenal dan pada umumnya digunakan untuk rekayasa genetik. Enzim tipe II dapat memotong antara kedua untaian simetris yang dikenal dengan pomotongan Blunt end (ujung buntu) atau mereka dipotong dengan sisi yang berbeda pada setiap unataian yang dikenal dengan ujung runcing (Cohesive end). Sekuens pengenalan pada enzim tipe II biasanya bersifat palindrome yaitu pembacaan dari arah yang sama pada masing-masing untai adalah sama.

Jumlah sekuens pengenalan menentukan kemungkinan pemotongan. Penemuan sekuens untuk sisi pengenalan empat basa lebih besar dibandingkan dengan enam basa pengenal. Jadi akan menyebabkan lebih sedikit, fragmen panjnag, enzim restriksi dengan enam atau lebih basa pengenalan.

Ketika dua sample DNA dipotong dengan enzim yang menyebabkan potongan sticky end, semua fragmen akan meninggalkan sekuen yang bergantungan. Ini akan membuat fragmen DNA dengan dua sumber dapat digabungkan bersama. Fragmen tersebut dihubungkan dengan DNA ligase, enzim yang sama yang menyatukan fragmen okazaki selama replikasi. Ligase yang umum digunakan adalah T4 Bacteriphage. Ligase mengkatalisis antara 3’-OH pada untai satu dengan 5’-PO­₄ pada DNA lainya. Ligase bekerja lebih efisien pada sticky end namaun juga bekerja pada blunt end.

https://www.youtube.com/watch?v=tcPgdR9_t64

https://www.youtube.com/watch?v=U2cKywEn6KY