IDENTIFICATION OF COPPER-INDUCIBLE GENES IN Pistia stratiotes

2.8 The use of living organism to monitor pollution 20
2.8.1 Bioindicator 21
2.8.2 Biomarker 22
2.9 Types of molecular biomarkers 25
2.9.1 Metallothioneins 25
2.9.2 Heat shock proteins 26
2.9.3 Phytochelatins (PCs) 27
3.0 MATERIALS AND METHODS 29
3.1 Materials 29
3.2 Culture media 30
3.2.1 Luria (LB) medium and LB agar 30
3.3 Stock solution 30
3.3.1 Antibiotic 32
3.4 Host strain and vectors 32
3.5 Preparation of glassware and plastic ware 34
3.5.1 Preparation of apparatus for RNA extraction 34
3.6 Maintenance of Pistia stratiotes 34
3.7 Determination of the rate of heavy metals uptake
by Pistia stratiotes 34
3.8 Isolation of total cellular RNA 35
3.8.1 Electrophoresis of RNA on denaturing agarose- 36
formaldehyde gel
3.9 DNase treatment of RNA 37
3.10 ACP-based RT-PCR 37
3.10.1 Reverse transcription 37
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