SIMPULAN DAN SARAN Simpulan
Transformasi konstruk gen chi ke dalam T. harzianum DT38 melalui A. tumefaciens strain AGL0 telah berhasil dilakukan dan transformannya memiliki
aktivitas kitinase 2 sampai 3 kali lebih tinggi dibandingkan non transforman. Transforman TH5 mempunyai aktivitas kitinase total paling tinggi sebesar 9.687 µM pNP/mg protein/jam dibanding non transforman T. harzianum DT38 dan T. harzianum DT41.
Saran
Berdasarkan hasil yang diperoleh pada penelitian ini, maka dapat dikemukakan beberapa saran untuk kelanjutan penelitian ini, yaitu :
1. penelitian untuk mempelajari aktivitas dan karakteristik enzim kitinase pada T. harzianum transforman dan tipe liarnya sebagai pembanding
2. penelitian untuk bioassay terhadap Ganoderma boniense secara in vitro dan in vivo
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Lampiran 1.
Komposisi beberapa pereaksi untuk isolasi DNA Larutan Pereaksi :
1. Larutan CTAB 10 % : NaCl 4,1 gram; CTAB 10 gram, dilarutkan dengan akuades hingga 100 ml.
2. Buffer Tris-HCl 1 M pH 8.0 : Tris-base 12,11 gram; HCl p.a 4,2 ml; akuades 80 ml, atur pH larutan hingga pH 8.0, tepatkan volume hingga 100 ml.
3. Larutan EDTA 0,5 M pH 8.0 : EDTA 18,61 gram; NaOH 2 gram; akuades 80 ml, atur pH hingga pH8.0, tepatkan volume hingga 100 ml.
4. NaCl 5 M : NaCl 29,22 gram, larutkan dengan akuades hingga 100 ml. 5. Ethanol 70 %
6. Isopropanol dingin 7. Ethanol absolut dingin
8. Chloroform : Isoamilalkohol (24 : 1)
9. Natrium Asetat 3 M pH 5.2 : CH3COONa 24,609 gram; akuades 80 ml, atur pH larutan hingga 5.2, larutkan dengan akuades hingga 100 ml.
10. Loading Buffer : Bromophenol blue 2,5 %; Sukrosa 40 % 11. Ethidium Bromida 1 % (w/v)
Buffer Ekstraksi Castillo (50 ml) : [ ] akhir
CTAB 10 % : 10 ml 2 %
EDTA 0,5 M pH 8.0 : 2 ml 20 mM
Tris-HCl 1 M pH 8.0 : 5 ml 100 mM
NaCl 5 M : 12,6 ml 1,26 M
Akuades steril : 20,4 ml Buffer TE (Tris-HCl : EDTA) :
Tris-HCl 1 M pH 8.0 : 1 ml EDTA 0,5 M pH 8.0 : 0,2 ml
Larutkan dengan akuades hingga 100 ml Buffer TBE 50X (500 ml) :
Tris-base : 27 gram Boric Acid : 13,75 gram EDTA 0,5 M pH 8.0 : 10 ml
