Ok, so I dont have much experience with microscopes, but i've been thinking about this for a long time.
If you could use a microscope, and say a small needle as your spore mover, Could you make a really dilute spore syringe and place a small drop on a slide from each strain, close enough so that they would be easy to join later, and use a small needle and some trial and error to get one and only one spore in each drop (granted after filddling with this long enough the drops would be really small) and then move each spore into the center in one very small water drop...then add a very small amount of sterilized LME/Dex LC solution on top of the spores, thus encouraging them to germinate.
Now when the monokaryotic mycelium from one spore meets the monokaryotic mycelium from the other spore, they will form dikaryotic mycelium, each cell of which would contain one nucleus from one strain and one nucleus from another strain...thus forming a new strain that would be a hybrid of the other two. One could then easily transfer that to a small liquid culture or agar petri and clean it up a bit since you might have contamination or just to grow it out to make enough health mycelium to inoculate a BRF jar...then from there you have spore prints and a new strain is born!
Now i realize that this would take many attempts to get right...and i also realize that one would have to do this in a sterile environment...then only thing that i dont have a full grasp on is the ability to move things that small around...it might be impossible It just seems like if it could work, then it would be much more reliable than the old way of letting two dikaryotic cultures grow into each other on a petri and hope that they exchange some genetic material through the cell walls...and even then you aren't guaranteed a complete hybrid... :'(
anyway...what do you think?
If you could use a microscope, and say a small needle as your spore mover, Could you make a really dilute spore syringe and place a small drop on a slide from each strain, close enough so that they would be easy to join later, and use a small needle and some trial and error to get one and only one spore in each drop (granted after filddling with this long enough the drops would be really small) and then move each spore into the center in one very small water drop...then add a very small amount of sterilized LME/Dex LC solution on top of the spores, thus encouraging them to germinate.
Now when the monokaryotic mycelium from one spore meets the monokaryotic mycelium from the other spore, they will form dikaryotic mycelium, each cell of which would contain one nucleus from one strain and one nucleus from another strain...thus forming a new strain that would be a hybrid of the other two. One could then easily transfer that to a small liquid culture or agar petri and clean it up a bit since you might have contamination or just to grow it out to make enough health mycelium to inoculate a BRF jar...then from there you have spore prints and a new strain is born!
Now i realize that this would take many attempts to get right...and i also realize that one would have to do this in a sterile environment...then only thing that i dont have a full grasp on is the ability to move things that small around...it might be impossible It just seems like if it could work, then it would be much more reliable than the old way of letting two dikaryotic cultures grow into each other on a petri and hope that they exchange some genetic material through the cell walls...and even then you aren't guaranteed a complete hybrid... :'(
anyway...what do you think?