lampiran 1. alat-alat penelitianmedia.unpad.ac.id/thesis/230110/2009/230110097019_l_5237.pdf ·...
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Lampiran 1. Alat-alat Penelitian
Hot plate
Cawan petri
Sentrifuse
Timbangan digital
Thermal cycler
Trays elektroforesis
Magnetic stirrer
Rak tabung reaksi
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Tips
Spatula
Botol Schott
Erlenmeyer
Tube PCR
Tube Eppendorf 1,5 ml
Autoclaf
UV-Transiluminator
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Tabung reaksi Shaker Inkubator
Rak Mikrotube Inkubator
Gelas ukur
Beaker glass
Kasa
Parafilm
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Sisir
Water bath
Vortex
Jangka sorong digital
Ose
L Glass
Laminar air flow
Sentrifuse
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Lampiran 2. Bahan-bahan Penelitian
(NH4)2SO4
NaCl
K2HPO4
MgSO4.7H2O
PDA
Agar
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Yeast extract
NaCl fisiologis 0,9 %
Akuades
Alkohol 70 %
Air fuchsin
Gentian violet
Objek glass
Pipet tetes
Lugol
Bunsen dan korek api
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EtBr
Primer ChiE-F dan ChiE-R
Loading dye Primer 16S rRNA
1 kb DNA ladder KAPA2G Fast Ready Master Mix
Isopropanol
EDTA
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Lysozyme
Tube falcon
Nuclei Lysis Solution
DNA Rehydration Solution
Protein Precipitation Solution RNase
Agarose
Nutrient broth
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Lampiran 3. Pembuatan Koloidal Kitin
Penjemuran cangkang udang Pemotongan cangkang udang
Persiapan alat dan bahan Proses pelarutan dalam HCL pekat
Koloidal kitin yang siap digunakan
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Lampiran 4. Proses Pengenceran Bakteri
Menuangkan agar Mengambil NaCl fisiologis 0,9 %
NaCl dicampur dengan bakteri Memasukan bakteri dan NaCl dalam agar
Meratakan menggunakan L Glass
Inkubasi hasil pengenceran bakteri
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Lampiran 5. Pewarnaan Gram Bakteri
Menyiapkan Pewarna Menyiapkan bunsen dan korek api
Mengambil satu ose bakteri Mengoleskan bakteri pada objek glass
Membilas gentian violet Membilas air fuchsin
Bakteri gram positif Bakteri gram negatif
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Lampiran 6. Kultur Saprolegnia sp.
Saprolegnia yang menyerang ikan Penuangan medium PDA
Menyiapkan peralatan kultur Saprolegnia disimpan di tengah medium
Saprolegnia diinkubasi 3 x 24 jam Pertumbuhan Saprolegnia selama 7 hari
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Lampiran 7. Isolasi DNA Genom Bakteri Asal Limbah Udang dan Kepiting
Kultur cair Proses sentrifugasi
Pelet DNA
Pengambilan supernatan
Memasukan lysozyme
Proses inkubasi dalam water bath
Memvortex larutan Memasukan isopropanol
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Lampiran 8. Analisis Bioinformatik Bioedit
1. Buka program Bioedit, kemudian buka file atau data yang akan diolah
menggunakan program bioedit.
2. Sorot (reverse) dilanjutkan dengan klik sekuen → Nucleic acid → Reverse
complement
3. Sorot (forward dan reverse) → Sequence → Pairwise Alignment → Align two
sequence (allow end to slide)
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4. Sorot (forward dan reverse) → Alignment → Create concensus Sequence
5. Klik basa pertama pada concensus → Edit → Select to end
6. Buka website http//ncbi.nlm.nih.gov → pilih BLAST → Nucleotide blast →
Masukan urutan sekuen → Pada database pilih 16S ribosomal RNA sequences
→ Klik BLAST
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Lampiran 9. Analisis Prediksi Struktur 3D Enzim Kitinase
1. Buka program swissmodel.espasy.org
2. Buka urutan asam amino hasil analisis BLASTX
3. Buka program SWISS MODEL klik my workspace
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4. Upload asam amino kedalam program my workspace
5. Tunggu sekitar 15 menit kemudian muncul