View Single Post
Old 05-20-2009, 11:28 PM   #10 (permalink)
Frankco
 
Join Date: Jan 2009
Posts: 91
BananaBucks : 6,666
Feedback: 0 / 0%
Said "Thanks" 4 Times
Was Thanked 37 Times in 18 Posts
Said "Welcome to Bananas" 0 Times
Default Re: what exactly is tissue culture?

Hi ArchAngeL01,

You have a pressure cooker, that's great!

The resident banana tissue culture expert, Gabe, put up a nice video on YouTube that shows how to trim the banana. You can watch it here.


As far as the part to use take a look at this page.
Micropropagation of Musa

As for the plant growth regulators, you will need these two, BAP and NAA. You can wait to get the NAA if you want. You won't need it until you are done multiplying the bananas. Depending on how many you want, you won't need it for at least a couple of months.

6-BENZYLAMINOPURINE SOLUTION or BAP (100 ml)
6-BENZYLAMINOPURINE SOLUTION (1.0 mg/mL) - Plant Growth Regulators - B130

NAPHTHALENEACETIC ACID SOLUTION NAA (100 ml)
NAPHTHALENEACETIC ACID SOLUTION (1.0 mg/mL) - Plant Growth Regulators - N605

You can get a smaller quantity of NAA from Carol's web site. You will not need as much of NAA as you will BAP. Carol sells it in 10 ml bottles. That’s enough for 20 liters of media as you will use .5ml per liter. Her shipping prices are better than Phytotech's too.
Home Tissue Culture* Group Catalog

You have to keep these plant growth regulars refrigerated. It is probably best to divide some in small plastic bottles and freeze it. They will last longer that way, about a year. Be careful that anyone who has access to your frig knows what they are. If there are children in your house, don’t keep them in the frig. It would be best to keep them in a cool, secure place and just buy fresh when you need more.

For Musa Basjoo you will need to use 5mg or 5 ml, ( 5mg=5ml with Phyototech’s BAP solution), per liter of media. This will give you the most multiplication. You should get at least 10+ little sprouts with the first explant. Cut each little sprout and put those in the same media and you should get 10+ from each one of those. The sprouts can be very small but each will grow. I got 16 from my first explant. The second round is proving to be just as good. So, on the second round you should get at least 10 x 10 or a hundred little explants in about 2 months. You should get better than this though.

You can get this agar to start out with. It’s $4.20 and will make a lot of media for you. Use 2 ¼ teaspoons per liter.
Amazon.com: Agar Powder - 2 Ounces: Health & Personal Care

Get some packets of MS nutrients. They are a $1 each. Each one will make 1 liter of media.
MURASHIGE & SKOOG (MS) BASAL SALT MIXTURE - Media, Basal Salts, & Vitamins - M524

Just pour the MS nutrients into 1 liter of distilled water. Add 4 teaspoons of sugar, 5 ml of BAP (use a 1 milliliter syringe to measure).

You will have to adjust the pH as it will be a little low. You want to get the pH close to 5.5 You can ballpark this with some pH papers. Just add the baking soda very slowly to raise the pH, a very tiny pinch at a time. Add 2 ¼ teaspoons of agar, heat to a boil and pour enough media into baby food or canning jars to cover ½ inch of the bottom and then put that in your pressure cooker for 15 minutes. You’re done with the media!

The web pages and video that I listed, I hope, will guide you the rest of the way.
Good luck and let me know how you do.
Frank
__________________
Frankco
Zone 5
N.W. Indiana
Frankco is offline   Reply With Quote Send A Private Message To Frankco
Said thanks: