Comment 1, 3, 4, 8 Analysing isozymes is simpler than doing polymerase chain reaction (PCR) followed by restriction fragment length polymorphism

(RFLP). Unquestionably, the technique RFLP is more accurate but for a very preliminary, large scale survey and for initial groupings of strains isolated from different localities the isozyme analysis is definitely a less costly (therefore can be done under a very small laboratory setup) and less time consuming method. The analysis of two isozymes can be done simultaneously in a dual vertical gel electrophoresis apparatus and within 6 hrs the whole profile can be generated and viewed. On the other hand DNA isolation, PCR and RFLP may take up to 20 hrs. Isozyme Preparation of enzyme extract- 20 min Gel preparation- 2 hrs Sample run- 2 hrs Gel staining Esterase- 1hr Catalase- 12 min SOD- 22 min RFLP DNA isolation 1.5 hr PCR 4 hrs RFLP- 12 hrs Agarose Gel Electrophoresis -3.5 hrs

Comment 2 - The missing reference I have included in the reference list. (Stock et al., 1998). Comment 6- I am not getting the question mam. Comment 7 I have made the desired correction. Please mam once go through it.