The healthy four week old sword suckers from Musa

The  healthy  four 
week  old  sword 
suckers  from  Musa
paradisiaca  Monthan cv.  Karibale weighing about one kilogram
uniformly sized were identified from elite plants of superior qualities such as
bunch weight of above 40 kg’s, more than ten number of hands in a bunch, more
than eighteen number of fingers in second hand from top and length of peduncle
more than 20 to 24 inches was selected and cut from the pseudostem 15 cm above the
base level, weighing 500 to 1500 g were collected from the banana fields of a
progressive farmer and used as starting material for micropropagation. These
suckers were transported to the Department of Biotechnology and  Bioinformatics,  Kuvempu 
University,  Shankaraghatta  and 
arranged  on  sand 
bed under shade and watered regularly.

Periodically Fungicides/ Bactericides are applied to these
suckers till they are taken for initiation process. The disinfection of suckers
was carried out thoroughly washed with tap water, soaked in a solution of 0.2%
Bavistin and 0.1% Streptocyclin for overnight. One whorl of leaf sheath was
removed and corm portion trimmed to a width of 1 inch x 1 inch and height about
2 inches, dipped in 0.001 M Teepol solution for an hour and finally rinsed
three to  four  time 
using  distilled  water 
to  remove   residual Teepol.  From such trimmed suckers shoot tip measuring
about 2.0 to 3.0 cm containing several sheathing leaf bases and axillary bud
along with underlying rhizome tissue were isolated.

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Surface sterilization:

The sucker cubes were transferred to the laminar airflow
chamber. Tissue  blocks  containing  
shoot-tips   and rhizomatous bases
were surface sterilized for six min in 70% 
ethanol,   0.2%   Mercuric 
Chloride  solution  for 
ten minutes  and  rinsed 
three  times  repeatedly 
with  sterile distilled water for
five minutes. One whorl of leaf sheath was removed, corm portion trimmed and
treated with 70% alcohol  for  six 
minutes,  0.1  % 
mercuric  chloride  for  ten
minutes  and  rinsed 
three  times  repeatedly 
with  sterile distilled water for
five minutes. Further, a whorl of leaf sheath was removed, corm portion was
trimmed, dipped in 1 % sodium hypochlorite for fifteen minutes and rinsed three
times repeatedly with sterile distilled water for five minutes.   Then the cut surface of the sucker tissue
was further trimmed and dipped in antibiotic solution containing Cifotaxime
0.1% and Gentamicin 0.05 % for 5 min. Finally, after the antibiotic treatment
the suckers were treated with Ascorbic acid (100mg/l) for 10 min to avoid
blackening of tissues due to phenolic exudation.