METHYL GREEN PYRONINE

                     METHYL GREEN PYRONINE

Solutions

1)    Pyronin Y - 500 mg

 Distilled Water-100ml

2)    Methylgreen-500 mg

 Distilled water-100 ml

                        3) Acetate Buffer (pH - 4.8)

Solution A

                        Sodium acetate                     -           136 gm.

                        Distilled water                        -           1000 ml

Solution B

                         Hydrochloric acid                 -          8.5 ml

                        Distilled water                        -           1000 ml

Composition of buffer

                                     Solution A                 -           50ml

                                    Solution B                  -           18ml.

           

                         Made up 250 ml with distilled water

Working solution

                              0.5% Methylgreen -        13ml

                             0.5% Pyronin Y               -        37 ml

                             Acetate buffer (pH 4.8)     -        50ml

                   Stock solutions are stored at 4°C.

Procedure

1)   Deparaffinise the section in xylene for 10 mts

2)   Bring the section to water through descending series of alcohol.

3)   Treat the slide with the working solution for 15-60 mts.

4)   Rinse quickly in distilled water and blot on non-fluffy filter paper.

5)   Flood slide with ethanol and leave for 3 minute.

6)   Flood with Xylene, leave until it clears and mount.

Result ·

                   Deoxyribonucleic acid (DNA)                            -        Green                                      Ribonucleic acid (RNA)                            -        Red