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Secotoid
Stranger



Registered: 04/21/19
Posts: 64
Last seen: 5 years, 8 months
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Next project
#25947931 - 04/21/19 05:20 PM (5 years, 8 months ago) |
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Having selected 3 fruits from my current tub based on early development, size and solidity of the stipe today is time to clone.
To keep myself entertained and because I had 4 qt of wheat spawn from the last batch I grabbed a spare shoebox and 2gal bucket which were laying around and spawned 2qt each to 3qt each of coir. They are currently covered with slitted cling film and standing by.

The clones will be made by tranfering interiol basal tissue to 3 plates per pin of meya and then at minimu. 1 transfer from each fruit will go on until clean.
The culture was a single transfer from Argentina MS plates to fresh ones selected arbitrarily from the largest homogenous sectors then to wheat.
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Secotoid
Stranger



Registered: 04/21/19
Posts: 64
Last seen: 5 years, 8 months
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Re: Next project [Re: Secotoid]
#25948482 - 04/22/19 12:43 AM (5 years, 8 months ago) |
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Gathering supplies while a half sleeve of 150mm ps pours of meya are cooling. Its almost cloning time 
Too bad i only have 2 half-sleeves left atm - it'll be down to reusing pp specimen cups again soon.
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Secotoid
Stranger



Registered: 04/21/19
Posts: 64
Last seen: 5 years, 8 months
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Re: Next project [Re: Secotoid]
#25948542 - 04/22/19 02:17 AM (5 years, 8 months ago) |
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While the agar was cooling I prepared the pins by slicing across half the thickness of the stems below the cap. This facilitates peeling it lengthwise when I am ready to remove interior tissue but keeps the cut well away from the clean basal tissue targeted. The lid was imediately returned to the tray they were stored in. I also wiped down all working and adjacent surfaces wtih 90% iso.
After the plates cooled I reopened the tray on my work area and peeled the first pin, being careful not to touch the exposed inner tissue and flame sterilized my scalpel blade. I cut three pieces for transfer to my plates, transferring each to it's plate immediately. After this I moved on the the next pin, reflamed my scalpel and onto the next set of plates until finished. I taped the plates closed with 13mm micropore tape and labled them. Since I had a spare pour and a spare pin (I like to have extra in case I contaminate something, drop it or likewise) I took a couple pictures. You all know the routine but I wanted to have more to show and didn't want everybody to think I was stopping to take pictures while doing clean work.



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