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http://indianaquariumhobbyist.com/community/ :: View topic - Aquatic Plants Tissue Culture
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Aquatic Plants Tissue Culture
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mandragoran
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PostPosted: Tue Aug 07, 2012 11:13 am Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Like  John  Travolta!   Guns
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garothmaan
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PostPosted: Tue Aug 07, 2012 11:35 am Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

                                                   
mandragoran  wrote  (View  Post):                
Like  John  Travolta!   Guns                

 
 Plant  fever..plant  fever.  Rock On   Innocent  
 
 I'll  try  a  coconut  tree  Surprised
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PRAVU
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PostPosted: Tue Aug 07, 2012 12:38 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Do  you  know  Garothmaan,  even  bamboo,  banana,  orchids,  everything  can  be  grown  in  tissue  culture?  Anyways,  good  going  sir,  lets  hope  for  the  best.  You  wont  find  fungal  contamination  in  day  1  or  2,  it  will  come  after  that.  So  observe  carefully.  Within  day  1  or  2,  there  will  be  bacterial  contamination,  if.......So  be  cautious.  And  best  of  luck.  And  for  the  identification,  the  bacterial  contamination  will  look  like  round  creamy  type,  and  the  fungal  will  be  cotton  looking.
 Thanks.
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garothmaan
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PostPosted: Tue Aug 07, 2012 1:23 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

                                                   
Madan  wrote  (View  Post):                

 
 Preparation  of  the  Media:
 
 1.  6-BAP  (  6-Benzylaminopurine,  a  Plant  Cytokinin  )  1  gram  of  powder  bought  from  ebay.com,  was  dissolved  in  200  ml  of  Dimethyl  sulfoxide
 and  stored  in  a  dark  container  in  the  fridge.
                 

 
 Approximately  How  much  time  here?  
 
 
                                                 
Madan  wrote  (View  Post):                

 2.  3-  IAA  (  3-  Indol  acetic  acid,  a  plant  Auxin)  1  gram  of  powder  bought  from  ebay.com,  was  dissolved  in  200  ml  Ethanol  and  stored  in  a  dark  container  in  the  fridge.
                 

 
 Approximately  How  much  time  here?  
 
 
                                                 
Madan  wrote  (View  Post):                

 3.  0.2  ml  of  each  of  the  above  solutions  was  added  to  1  liter  of  boiled  and  cooled  water  to  get  a  concentration  of  1mg/l  of  each  hormone.
                 

 
 2  seperate  container  right  (  each  means)  dont  mind  my  silly  question  pls  Smile  
 
 
                                                 
Madan  wrote  (View  Post):                

 4g.  A  little  Vinegar  was  added  to  reduce  the  pH,  pH  checked  and  found  to  be  about  5.0
                 

 
 White  Vinegar  right?
 
 
                                                 
Madan  wrote  (View  Post):                

 4f.  Sodium  bicarbonate  powder  was  then  added  to  the  media  solution  and  pH  adjusted  to  5.8  (approx).
                 

 
 what  was  used  to  meassure?
 
 
                                                 
Madan  wrote  (View  Post):                

 4f.  The  bottle  and  media  solution  +  agar  was  then  microwaved  and  cooled.
                 

 
 Approximately  how  much  time?
 
 
                                                 
Madan  wrote  (View  Post):                

 5b.  The  ex-plants  were  then  transferred  to  the  tissue  culture  bottle  with  the  cooled  and  hardened  media.  Bottles  closed  and  labelled.
                 

 
 Tight  lids  or  just  enough  to  allow  any  air?
 
 
 @  PRAVU  Saheb,  I  know,  I  read  the  link  kitchen  culture,  very  impressive  and  enlightening.  If  only  all  those  rare  herbs  from  himalayas  could  be  cultured  for  diseases  cure.  Thumb Up
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PRAVU
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PostPosted: Tue Aug 07, 2012 1:28 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Yes  Garothmaan  ji,  a  lot  of  works  on  the  herbs  are  going  in  different  labs.  I  have  seen  some  of  them.  They  are  really  great.  Thumb Up  .
 Induction  with  different  hormones  are  really  a  good  thing  to  watch,  and  slowly  but  surely  there  effects  can  be  observed,  thats  a  great  experience.
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garothmaan
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PostPosted: Tue Aug 07, 2012 1:40 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

                                                   
PRAVU  wrote  (View  Post):                
Yes  Garothmaan  ji,  a  lot  of  works  on  the  herbs  are  going  in  different  labs.  I  have  seen  some  of  them.  They  are  really  great.  Thumb Up  .
 Induction  with  different  hormones  are  really  a  good  thing  to  watch,  and  slowly  but  surely  there  effects  can  be  observed,  thats  a  great  experience.                

 
 THANK  YOU  FOR  DOING  ALMIGHTY  GOD'S  WORK-  I  pray  that  you  and  people  like  you  shall  attain  success  in  your  good  work  to  help  mankind  and  make  our  country  proud  Bow  Bow  Bow
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PRAVU
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PostPosted: Tue Aug 07, 2012 2:43 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Thanks,  I  am  planning  to  do  something  good  in  future.  anyways,  its  all  our  hard  work  that  will  make  our  country  proud.  
 Thanks.  Best  of  luck.
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Madan
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PostPosted: Tue Aug 07, 2012 5:25 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 garothmaan
 
 1.  30  secs
 2.  60  secs
 3.  one  container
 4.White
 5.nothing
 6.  60  secs  and  forever
 7.  Tight
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PRAVU
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PostPosted: Tue Aug 07, 2012 7:27 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Tight  is  good  to  avoid  contamination,  but  you  need  to  check  out  whether  they  are  growing  or  dying  as  they  nedd  aerobic  conditions  as  they  are  higher  eukaryotes.
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Madan
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PostPosted: Tue Aug 07, 2012 9:51 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 48  hrs  no  bacteria  in  my  samples.
 Fingers  crossed  !
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PRAVU
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PostPosted: Tue Aug 07, 2012 10:14 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Good  going  sir  ji.  What  hormone  did  you  add?  I  mean  for  the  callus  culture?  What  are  you  expecting?
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Madan
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PostPosted: Wed Aug 08, 2012 10:53 am Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Prabuddha  we  added,
 
 6-Benzylaminopurine  a  cytokinin  at  1mg/l  concentration  of  media,
 along  with  3-Indol  acetic  acid  an  Auxin  at  the  same  concentration.
 
 Your  suggestion  of  liquid  media  might  just  work  for  aquatics,  as  I  just  now  got  some  info
 from  a  person  who  has  succedeed  with  Glossostigma  elatinoides  and  Hemianthus  callitrichoides
 at  home.
 His  media  is  just  about  solid  in  the  grow  out  stage,  he  says  it  is  very  loosely  solid,  consistency  much  thinner  than  silicon  just  out  of  the  tube.
 
 He  has  promised  me  pictures,  hope  to  get  them  soon.
 
 Thanks  for  the  tip  !
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Cherry
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PostPosted: Wed Aug 08, 2012 2:17 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Hello  Madan  Ji,
 
 M  so  happy  to  hear  that  48hrs  have  passed  with  no  contamination.M  sure  the  culture  will  come  out  great.
 Excited  to  see  the  results.
 But,  sad  couldnt  be  a  part  of  the  ordeal....extremely  held  up  with  a  cousins  marriage,then  going  home  to  visit  my  parents.
 kindly  keep  uploading  photos  of  the  progress,will  be  glad  to  see  them.
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Ravabi
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PostPosted: Wed Aug 08, 2012 2:23 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 That's  great  news   Rock On  .....Are  these  the  second  batch  which  you  did  after  we  left
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Madan
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PostPosted: Wed Aug 08, 2012 2:24 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Yes.
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