Bodgy DIY Tissue Culture Method

Bodgy DIY tissue culture method

This tissue culture media can be used for experiments with cuttings clones, or for seed germination.
This is not how the professionals do it, but I figure you folks may still be interested in my low chemical, bodgy DIY method, that does the job well, and costs bugger all.
I have used it for experiments on some super hard to grow species and it gave me great results and was fun, so I figured I should share the info.

Ingredients

  • 15g of pure fine agar powder.
  • 20g of white cane sugar.
  • 1.25g of pure thiamine. This is Vitamin B1. You can buy it pure online, or just crush 5 tablets 250mg tablets from the chemist or supermarket. The added talc, fillers and gelling agent(agar) in the tablets have no noticeable effect on results.
  • 40g of ripe banana. Half a small fully ripe, but not at all brown one.
  • 2 medium fully ripe tomatoes or a rough approximation.
  • 900ml of rainwater or failing that tap water. Use distilled water if your tap water smells like bleach, sulfur, or is above pH 7, but keep in mind the lack of minerals will have a negative effect.

Sterilization advice.
All of the following equipment is meant to be dipped in bleach, rubbing alcohol, or hydrogen peroxide. Failing this it is meant to be boiled in a pressure cooker for at least 30 minutes. “Sterile” is the goal but even though I do none of that…

I just wash really really really well in rainwater, and make believing like I am some sort of M.A.S.H. surgeon.
I sit all my tools on a large stainless steel tray that is also super clean.
I DO NOT EVER PUT A TOOL DOWN, THEN LATER PICK IT UP AND REUSE IT!
If I finish using a whisk or tongs for example, I then dump it in the sink or a bucket to be washed later(not touching anything else with my super clean hands in the process).
Later if I need a whisk or tongs again, I just grab another clean one from the pre-prepared tray.
You can not be too organized, or too clean!

Tools and equipment.

  • 1x large tray or plate big enough to sit all your super clean tools on.
  • 1x large clean stainless steel pot (NOT aluminium or it will react with the acids).
  • 1x large clean stainless steel sieve.
  • 2x tongs, just in case I need to touch something “dirty”.
  • 2x whisks (1 is a spare in case I bump something “dirty” or drop it or whatever).
  • Disposable gloves and face mask is well worth considering too. All the experts use them, folks far wiser and more experienced than me…

Growing vessels.
I use small cheap blood sample tubes(15ml-30ml-50ml) for cuttings, and I use short round takeaway 100ml sauce tubs for seeds.
These should be laid out on trays or in racks ready to be filled as the liquid growing medium is cooling.
Again these should be sterilized, but again I never bother…

Method.

  1. Fully whip, blend, or food processor the 20g of white cane sugar, 1.25 g of pure thiamine(or 5x 250mg B1 tablets), 40grams banana(~half), 2 medium fully ripe tomatoes and 900ml rainwater.
    Depending on your maceration tool of choice it may be best to add the water a little bit at a time instead of all at once.
  2. Sieve this several times then whisk in the 15grams of agar powder and place in the stainless steel pot on the stove.(no lid on the pot!)
  3. Bring to the boil while whisking for 10-15minutes. Use a whisk to stir rapidly and continuously, so the sugars don’t caramelize. Burnt sugar has lost most of its energy, and the plants need that energy. Plus sugar is a preservative and it decreases bacteria and fungal issues.
  4. Kill the heat and move to the side to cool without a lid! (condensation water drips and causes irregularities in consistency and temperature).
  5. Once it completely stops bubbling and is cooled to sensible levels that won’t melt your plastic or burn you, but is still “hot” pour it into the moulds only filling halfway maximum. Put on lids, but do not “click” them sealed. Just leave sitting on top to cool fully and set. It is not jelly(gelatine), and it does not need to go in the fridge, normal room temperature is fine.
  6. Once fully cooled and 100% set, add your very very clean seeds or cuttings and click or screw shut your lids and place them in an area with lots of cool light. I just use a table on the balcony, but some folks use LED or Fluorescent lights @12hrs -18hrs light a day).

The seeds or cuttings should also be dipped in bleach or peroxide before adding, but I never bother bleaching or sterilizing any of my equipment and I still get a very low level of losses from contamination, because I am super careful.
I wash everything very well in running rainwater. I shake all tools dry and sit in the sun for on clean dry stainless steel trays for 10-20mins before use.

The key is, I am super obsessively clean and I never ever touch or put anything down once I start. Don’t lean over anything, or cough, or scratch your nose and definitely don’t adjust your jocks!

You can also try adding various concentrations of GA3, or using very weak, very well sieved Smoke Water for improved seed germination.
You can add various concentrations of Indole-3-butyric Acid (IBA) “Rooting powder” for improved root development in cuttings.
You can try making up and mixing in some using my Willow water recipe which has a mix of both Indolebutyric acid (IBA) and Salicylic acid (SA) for root development and immune response.

Have fun folks!