Home | Community | Message Board


Mycohaus
Please support our sponsors.

Mushrooms, Mycology and Psychedelics >> Advanced Mycology

Welcome to the Shroomery Message Board! You are experiencing a small sample of what the site has to offer. Please login or register to post messages and view our exclusive members-only content. You'll gain access to additional forums, file attachments, board customizations, encrypted private messages, and much more!

Amazon Shop for: Agar, Petri Dish, Potato Dextrose

Jump to first unread post. Pages: 1
OfflineQuincunx
Whisper
Male User Gallery

Registered: 02/21/03
Posts: 72
Loc: Slovenia
Last seen: 1 year, 7 months
tissue cloning & DNA degeneration with cubes?
    #2276613 - 01/27/04 04:45 AM (13 years, 3 months ago)

When using fruitbody tissue to isolate a cubensis substrain and then store them in agar slants in a fridge - how long until one can expect DNA breakdown if fresh slants are used every 6 months and re-inoculated with previous colonized slants?

Thanx in advance!


Post Extras: Print Post  Remind Me! Notify Moderator
Offlinezeronio
Stranger
Male

Registered: 10/16/01
Posts: 2,349
Loc: Slovenia
Last seen: 7 months, 20 days
Re: tissue cloning & DNA degeneration with cubes? [Re: Quincunx]
    #2276740 - 01/27/04 06:36 AM (13 years, 3 months ago)

Very long. Cubensis does not senescence (degenerate) as fast as some other species. I have succesfully fruited them from a P10 petri, which means that mycellium grew over 10 petris after cloning. In your case that would mean about 5 years. If you use the distilled water storage in which you can store mycellium for several years then you can forget about senescence.


Post Extras: Print Post  Remind Me! Notify Moderator
OfflineQuincunx
Whisper
Male User Gallery

Registered: 02/21/03
Posts: 72
Loc: Slovenia
Last seen: 1 year, 7 months
Re: tissue cloning & DNA degeneration with cubes? [Re: zeronio]
    #2277474 - 01/27/04 01:38 PM (13 years, 3 months ago)

Quote:

zeronio said:
If you use the distilled water storage in which you can store mycellium for several years then you can forget about senescence.



Ok. Great. I'll go rather with this method. Seems more fullproof and long lasting. As i read in the FAQs, sealed test tubes can be thus even stored at room's temperature this way (exellent :grin:).
But... (from http://www.shroomery.org/index/par/23543 ) :
Quote:

Viability deteriorated rapidly after 2 years storage and 42% (21/50) of the isolates were dead at 5 years. Growth may sometimes occur during storage in water. This will be reduced if the spores or hyphae are removed from the surface of agar media and no medium is transferred.



How do you guys do that? I mean - separate nutrients (agar) from mycellium? Is there any TEK avaiable?
I have access to a lab centrifuge machine, but cube mycellium is sensitive to bruising, so how should i set the centrifuge? Would 2000-2800 RPMs be too much? How many washings is adequate?
I could ask there, but i don't dare. :shake:


Post Extras: Print Post  Remind Me! Notify Moderator
Offlinezeronio
Stranger
Male

Registered: 10/16/01
Posts: 2,349
Loc: Slovenia
Last seen: 7 months, 20 days
Re: tissue cloning & DNA degeneration with cubes? [Re: Quincunx]
    #2279685 - 01/28/04 03:24 AM (13 years, 3 months ago)

Most species grow only on surface of agar so it's not hard to peel small chunks of mycelium off it with tweezers or scalpel. If you use several vials for each culture then you have good chances of maintaining it, even if some agar gets inside.
I plan to revive all cultures once a year and so far I had very good results.

I wouldn't use the centrifuge - it would be very hard to maintain the sterility which is very important in this case.


Post Extras: Print Post  Remind Me! Notify Moderator
OfflineQuincunx
Whisper
Male User Gallery

Registered: 02/21/03
Posts: 72
Loc: Slovenia
Last seen: 1 year, 7 months
Re: tissue cloning & DNA degeneration with cubes? [Re: zeronio]
    #2279968 - 01/28/04 08:51 AM (13 years, 3 months ago)

Mission accomplished :thumbup:. Thanx Zeronio.

I used spatula to remove mycellium from agar surface, then i used Ependorf tubes for storage in dH2O - they seal great, absolutely no air inside and they are small and neat. But it took me almost 2 and half hrs to fill 20 of them. :smirk:



Post Extras: Print Post  Remind Me! Notify Moderator
OfflineSuntzu
Geek
 User Gallery

Registered: 10/15/99
Posts: 1,394
Last seen: 19 days, 15 hours
Re: tissue cloning & DNA degeneration with cubes? [Re: Quincunx]
    #2280203 - 01/28/04 11:34 AM (13 years, 3 months ago)

How adherent the mycelium is to the agar depends a lot on the particular species [defined here :wink: ] and the type of agar used.  I have worked with agar that mycelium very much 'burrows into', but the standards [e.g. PDA] do not seem to do so. 
The tool of choice I prefer for making dH2O suspensions is a sterilized swab.  Rubbing it over the culture picks up the fragments rather effeciently with a minimum of agar material, and the swab can be immersed in the destination water and twisted, nicely distributing the fragments in solution.  I really can't say enough good about storing cultures in dH2O; tiny aliquots can be taken at any point [if you have access to a pipet, all to the good] for direct innoculation of grain [or agar]. 
Those little eppendorfs will serve you well;  You can also easily get away with using 15 mL or larger tubes, a lot less work. 

Nice lab bench, BTW :wink:


Post Extras: Print Post  Remind Me! Notify Moderator
Jump to top. Pages: 1

Amazon Shop for: Agar, Petri Dish, Potato Dextrose

Mushrooms, Mycology and Psychedelics >> Advanced Mycology

Similar ThreadsPosterViewsRepliesLast post
* Sterile Tissue Cloning Procedure BigBlackNinja 1,467 9 03/20/03 11:54 AM
by Anonymous
* Dry tissue cloning/spore extraction ??? Bi0TeK 1,525 2 06/25/03 12:45 PM
by Bi0TeK
* tissue clone of Psilocybe atlantis lafeeverte 1,377 5 12/09/04 01:15 AM
by Koala Koolio
* Three Questions about Senescence FugueRider 767 2 03/29/09 10:11 PM
by FugueRider
* Cardboard cloning tek?
( 1 2 all )
ink 10,992 31 02/15/16 12:43 AM
by Eywa_devotee
* tissue and grain
( 1 2 all )
La_Magdalene 2,661 32 02/01/03 03:42 PM
by La_Magdalene
* 5 questions on cloning blackout 2,018 6 08/22/01 01:16 AM
by jonnyshaggs420
* Transfers vs Cloning Mycorial 876 7 05/12/09 09:04 PM
by Mycorial

Extra information
You cannot start new topics / You cannot reply to topics
HTML is disabled / BBCode is enabled
Moderator: Prisoner#1, RogerRabbit, EvilMushroom666
1,350 topic views. 0 members, 3 guests and 1 web crawlers are browsing this forum.
[ Toggle Favorite | Print Topic | Stats ]
Search this thread:
The Spore Depot
Please support our sponsors.

Copyright 1997-2017 Mind Media. Some rights reserved.

Generated in 0.046 seconds spending 0.001 seconds on 14 queries.