Page 5 of 8

Re: Plant Tissue Culture

Posted: Fri Oct 05, 2012 11:57 pm
by maaz
Hello guys here is one of the Media list they just send me today and i need you help regarding this which one better too use..i am uploading this pdf converted to excel file here for all..In Pdf lots of products for Tissue culture i am just pasting for MURASHIGE & SKOOG (MS)



MURASHIGE & SKOOG MEDIUM (With Vitamins & Folic Acid) 5x1 Ltr 1x10 Ltr 1x50 Ltr 350 450 1660
MURASHIGE & SKOOG MEDIUM (With Vitamins & MES Buffer) 5x1 Ltr 1x10 Ltr 1x50 Ltr 375 450 1600
MURASHIGE & SKOOG MEDIUM (With Vitamins, W/O Sucrose, Agar & Calcium Chloride) 5x1 Ltr 1x10 Ltr 1x50 Ltr 325 360 1450
MURASHIGE & SKOOG MEDIUM (With Vitamins & Calcium Chloride W/O Sucrose, Agar) 5x1 Ltr 1x10 Ltr 1x50 Ltr 325 360 1450
MURASHIGE & SKOOG MEDIUM (With Sucrose, & Vitamins, W/O Agar & Calcium Chloride) 5x1 Ltr 1x10 Ltr 1x50 Ltr 325 360 1450
MURASHIGE & SKOOG MEDIUM (With Sucrose, Agar & Vitamins,W/O Calcium Chloride, IAA & Kinetin) 5x1 Ltr 1x10 Ltr 1x50 Ltr 440 495 1820
MURASHIGE & SKOOG MEDIUM (S.B) (W/O Vitamins and with Calcium Chloride) 5x1 Ltr 1x10 Ltr 1x50 Ltr 220 250 990
MURASHIGE & SKOOG MEDIUM (S.B) (W/O Vitamins & Calcium Chloride) 5x1 Ltr 1x10 Ltr 1x50 Ltr 220 250 990
MURASHIGE & SKOOG MEDIUM (S.B) (W/O Calcium Chloride & Vitamins) 5x1 Ltr 1x10 Ltr 1x50 Ltr 220 250 990
MURASHIGE & SKOOG MEDIUM MODIFIED (SB)With 1/2 Macronutrient 5x1 Ltr 1x10 Ltr 1x50 Ltr 220 255 990
MURASHIGE & SKOOG MEDIUM MODIFIED (SB) W/O. NH4NO3 5x1 Ltr 1x10 Ltr 1x50 Ltr 220 250 990
MURASHIGE & SKOOG MEDIUM MODIFIED (SB) ½ x NH4NO3 and KNO3 5x1 Ltr 1x10 Ltr 1x50 Ltr 220 250 990
MURASHIGE & SKOOG MEDIUM MODIFIED (SB) With Micro and Macro nutrient W/O. Ammonium Salt 5x1 Ltr 1x10 Ltr 1x50 Ltr 220 250 990
MURASHIGE & SKOOG MEDIUM (SB) With minimal organics (MSMO) 5x1 Ltr 1x10 Ltr 1x50 Ltr 220 250 990
MURASHIGE & SKOOG MEDIUM (SB) With Gamborg’s Vitamins 5x1 Ltr 1x10 Ltr 1x50 Ltr 295 360 1265
MURASHIGE & SKOOG MEDIUM Vitamin Powder (1000x) 100 ml 385

Re: Plant Tissue Culture

Posted: Sat Oct 06, 2012 1:28 pm
by SCORPIO
Maaz, Keep it up.

Re: Plant Tissue Culture

Posted: Sat Oct 06, 2012 1:37 pm
by prajjwal
Maaz mail me the pdf at prajjwalg@gmail.com please

Re: Plant Tissue Culture

Posted: Sat Oct 06, 2012 1:45 pm
by parthapratim22
Maaz Bhai in this thread pdf is not allowed, kindly contact Rajivji.

Rajivji please help to upload the file. Will send you in your personal mail.

Re: Plant Tissue Culture

Posted: Sat Oct 06, 2012 2:04 pm
by prajjwal
maaz wrote:MURASHIGE & SKOOG MEDIUM (With Sucrose, Agar & Vitamins,W/O Calcium Chloride, IAA & Kinetin) 5x1 Ltr 1x10 Ltr 1x50 Ltr 440 495 1820
It will do IMO...get Frankco's opinion too...

Re: Plant Tissue Culture

Posted: Sat Oct 06, 2012 3:11 pm
by maaz
prajjwal wrote:Maaz mail me the pdf at prajjwalg@gmail.com please

email send...u can check buddy...yes i am also looking for Frank opinion for this..

Re: Plant Tissue Culture

Posted: Sat Oct 06, 2012 3:40 pm
by SCORPIO
PDF File sent by Partha
Plant Tissue Culture Media_009.pdf

Re: Plant Tissue Culture

Posted: Sat Oct 06, 2012 3:46 pm
by maaz
oohhh...thanks Rajivji thanks of uploading..

Re: Plant Tissue Culture

Posted: Sat Oct 06, 2012 8:32 pm
by Frankco
Hello everyone,

Thank you for all the kind words.

I have been researching media formulations for aquatic plants to try and get a general idea on the media formulations. The tricky part with plant tissue culture is that each species might require different media.

Generally, I found that MS (Murashige & Skoog Basil Medium) plus Gaborg’s vitamins with 100 milligrams of Inositol seems to be a common type of media for aquatic plants. Again each species may require something different but this is what I would start with. Even if some aquatic plants need different media I would expect this media to produce growth.

There are many types of media on Titan’s website. Do not let all of the different types of media overwhelm you. Many are the same but with slight differences in ingredients optimized for different types of plants. That does not mean that plain MS media will not work too along with the modified MS version. The modified version will just work a little better. Sometimes MS will not work and you have to use another type media. Google will help with selecting the correct media. Search words such as the botanical name of the plant and “in vitro” or micropropagation or regeneration.

MS Media was developed specifically for Tobacco plants. The formula was optimized to produce the maximum growth in tobacco. To optimize growth in other plants different amounts of the ingredients may be required. To it make easier for tissue culture labs, so they do not have to measure out each ingredient separately, suppliers of the media make the adjustments, pack it up and label the media accordingly. You will see MS modified media or MS without calcium chloride etc.

MS media was found to work with a large variety of plants. It is the most common media used in plant tissue culture. That said, many plants cannot tolerate the high amount of ammonium nitrate in MS, especially some woody plants and definitely carnivorous plants. Some plants need additions or adjustments to chemicals in the mixture. Different types of media have been developed for different types of plants. Hence the variety of media listed on Titan’s site.

You can see the list of ingredient in product number TP 025 on Titan’s site. I would go with this media for aquatic plants.
http://www.titanmedia.in/plant-tissue-c ... -2-12.html

It is very difficult to create your own media. You have to have a very sensitive scale. You have to mix each chemical independently for a stock solution and make sure it does not come out of suspension. If it does then you might have make it all over again, increasing costs. You would take a small amount of each chemical from the stock solutions and mix those to make your final media for the plant. Mistakes are easy to do at every step. One miss measured ingredient can kill you plants.

I calculated the cost of making MS from buying chemicals off of eBay. It came to about $212.56 US dollars for to make 453 liters.
453 liters was the minimum amount I could make from buying each of chemicals separately. It seems like a cheaper way to go but it is very difficult. Buying premixed chemicals lessens the chance of mistakes and makes life easier.

When you first starting out you want the easiest path you can take as trying to guess what went wrong when a plant dies is more difficult when the procedure is complicated by mixing your chemicals. You buy smaller amounts and not have to worry about storing all of the different chemicals.

Also you will when you start out using a packet that makes 1 liter of media you will looking for a way to store the extra media. It should be refrigerated and you can store it for about 4 weeks if the bottle you put it in is clean. Do not store media that has sugar (sucrose) in it. Leave out until you are ready to use it.

One quick thing about agar. 7.5 grams per liter is the average amount for plants. This can vary with the type plant. To save costs a paper coffee filter can be crumpled up and placed in a jar to support the plant. This will not work for all plants but would work quite well for aquatic plants. I’ll include a picture of bamboo I have growing using a coffee filter using liquid media without agar. It is just a way to save money on the cost of agar.

Image

Very fun post. I wish you all luck and I will try to help you in anyway that I can.
Frank

Re: Plant Tissue Culture

Posted: Sat Oct 06, 2012 9:42 pm
by prajjwal
First of all, thanks a lot to Mr. Frank for sharing his experience and knowledge with us. I think your help will surely make us successful in this project.

Guys, we should make list of plants which we should try to Micropropagate and easy or hard, will be judge by Mr. Franc. It will be easy for even for him to designing a media for that plant. Accordingly we all (3 of us atlist) will start with it and wherever we will face any problem, we will wright in this post.

or----------

What about starting with the most easiest and fast growing plant (even terrestrial) to test our skills and setup. Fighting with contamination and understanding the minimum requirements of a plant to grow in a complete artificial media. Then we can shift to most easiest Aquatic plants, like Anubias, Ferns, Grass, Cryptocorynes etc etc....

What you people think??

Regards,
Prajjwal Ghosh