MS (Murashige and Skoog, 1962) media supplemented with
different and various combinations of plant growth regulators
were used for shoot-tip culture. Myo-inositol and sucrose were
added freshly. Plant growth regulators from stock solutions
were added as per requirement. The pH of the solution was
adjusted to 5.8 using 1N NaOH or 1N HCl and the final volume
made up. After making up the final volume, the solution was heated and agar @ 0.8 % was dissolved to make the media
semi-solid. While heating, the media was stirred frequently
with a glass rod or shook frequently for uniform heat
distribution and to avoid frothing and boiling. After the agar
was dissolved completely, the media was distributed into testtubes
(150 x 25 mm borosil tubes) or conical flasks (100,150,
250 ml volume). The tubes and flasks were plugged tightly
with non-absorbent cotton wrapped with cheesecloth. The
culture vessels containing media were sterilized by autoclaving
at 15 p.s.i(121°C) for 20 minutes. After autoclave sterilization
they were cooled at room temperature and then transferred to
the inoculation room.