FlytrapCare Carnivorous Plant Forums

Hi,

I'm new here but I making TC for around 50-60 jars.They contamination 98-99% because I have no experience and technics especially I don't have pressure cooker, I will buy some soon. Now I learn TC myself and understand it, last 2 jars are success they are tomato and carex grass
Now I will experiment with tyndallization method before buy new pressure cooker. (http://en.wikipedia.org/wiki/Tyndallization)
I will update result of them again

I have one question when put jar in pressure cooker. Do you slightly open lid for air ventilation in-out the jar ?

Thank you very much and sorry if my English not good.

Hello and Welcome to FlyTrapCare. It's good too hear that you have a bit of success with your TC. It's always an experiment but don't worry, you will get the hang of it soon.

Now to answer your question, you don't need to slightly open the jars. Just leave it closed.

hb

hackerberry already answered your question. I just wanted to say hello and welcome. And to encourage you not to give up on TC. The learning curve is steep and there is a lot of initial failure, but once you develop your technique, it's fun!

HB already answered

Hello an many welcomes!

Thanks everybody.
If closed the jar,Pressure in jar will not higher ,right ? It still less than 15 PSI.
Why it can sterilyzation ?

natapongw wrote:If closed the jar,Pressure in jar will not higher ,right ? It still less than 15 PSI.

No, that's not correct. The jars aren't air tight. The pressure is the same inside the jar.

We-'Found' At PLANTECH IN-The Mid-Nineties ... that The 'Critical'-Temperature for The 'Hot'-BLEACH Technique Lay-around The 53oC Mark ... Which-IS 'Not'-Far From The Scoulding-Temperature of ~60oC of-Most Average House-House HOT-Taps so-to-Speak. >(*U^)<

"I"-Can't Off the-Top-of My-Head Remember 'What' The REMELT-Temperature of Agar IS ... but the Concept of Placing-Your Jars IN-The-Household Laundry-Trough THREE-Days IN-Sucession & Doing Nothing-Else IS VERY-'Appealing' INDEED "I"-Must Admit!!! >(*U^)<

PERHAPS Some Intellectual-Compromise Between Tyndellization & The-Above Can-be 'Made' ... if-R Suitable Sterilant Can-be Found so-to-Speak!!!??? >(*~*)< / >(*U^)<

Also CHEAP Agar from most Asian-(Supply)-Shops / Houses R Good-enough for Plant-Tissue Culture Purposes &-R Extremely Versitile as-Well-as Cost-Effective.

http://www.flytrapcare.com/phpBB3/first ... 2-105.html:

FROM Last-Night

"My-BRAIN Went-'Snap' about An-Hour-ago. >(*U^)<

'Whoever' Gave-that Link to TYDALLIZATION Has-Probably 'Sucked'-Me-IN to-Trying SULPHUR-Powder as-R Long-Term Sterilant In The-Laundry Trough Water-Bath so-to-Speak >(*~*)< You-'mean dude'!!! >(*U^)<

(1) Add-Quantity (to-Be 'Determined') of Sulphur-Powder to-'Molten'-AGAR Medium.

(2) Allow-to-Set.

(3) Plant Explant-'Cuttings' BELOW 'SOFT'-Agar Medium (Probably from-15-Year-Old 'Memory': 1.25g/Litre).

(4) TYNDALLIZED in ~60c Laundry-Trough 'Water'-Bath THREE-Days IN-Succession. [as the Jars Come-UP to Temp ... The-Medium 'Should' Rhoughly-Hit The MAGIC 53oC ... that-Causes NO-(Appreciable)-Damage to Most Large-Size Explants (Especially the-Phenolically-Ones).

(5) See-What 'HAPPENS'!!!??? >(*~*)< / >(*U^)<

Certainly Fits The KISS-Principles Outlined at the Header of This Topic ... but WILL-It 'Work'!!!??? >(*~*)<"

****

Well Since Going-to-Bed Last-Night "I"ve Dug-UP Some Sachets of CHEAP Chinese Agar-Powder "I"-'Knew' "I"-Had IN-R-Drawer Somewhere:





From The-LABEL of The 'Dusting'-Sulphur "I"-Have IN-The-Shed ... "I"ve-'Noticed' that It-Says that it's 'Not'-ONLY R-FUNGICIDE ... But 'Also' R-MITICIDE!!! >(*~*)< R-Common-Problem of Tissue-Culture Spore-Vector-Transfer!!! >(*U^)<

If-You've 'Ever' Played-around with Sulphur You'll-'Know' that It 'STICKS'-like Buggery ... but "I"-'Think' Our First-Step Is-to Run R Standard Control Against Sugar With-&-Without Sulphur & With-&-Without 60oC Household 'Hot'-Tap / Laundary-Bath TYNDALLIZATION ... 'Just'-to-See if-There Is 'Some'-Merit IN-The-Process and Whether-or-'Not' It's Worth-Proceeding so-to-Speak!!!??? >(*~*)< / >(*U^)<

Well ... "I"m-Beginning to Get-'Bach' INTO The-Swing-of-Things.

Made-UP R-Batch of 1cc / 600Mls Just-to-'See' if the-Old Agar 'Worked' so-to-speak. It's-R-Little-ON The-'Soft'-Side ... though should Firm-UP R-Bit Over The-Next 24-Hours as-Agar Does ... but Perhaps 2ccs made-UP to 600mls Would-be 'Ideal' for Plunging Large Explant-Material INTO Intimate-Contact with The-Medium!!!???



'Had'-R-(Playaround)-GO with Sulphur & Biodegradable Household Surfactant. If-One 'Assumes' Rhoughly 60-Drops Per 1ml Then This 1-in-3600 Dilution (IE one-Drop IN 600mls) Appears Quite-'Adequate' &-'Should' Suffice If-Require-or-'Needed'.



Initial-Suggestion ... 'Just'-from The-Above Work Is-that Surfactant is-Likely 'Not'-Required and-That The-Sulphur is-Probably 'Best' Mixed-IN with ~300-Stirs as-soon-as the Agar cools but 'Just'-before it-Sets / 'Gels' so-to-Speak.

TOMORROW "I"ll TROLL for Cheap-Jars / Vessels &-Perhaps Buy-some SEAWEED-Extract that-Is 'Supposedly' Chock-FULL of 'Natural'-CYTOKININS &-hopefully Adhere-to The KISS-Principle before "I"-Really Get Stuck-IN to The-Job-@-Hand: CHEAP Set-&-Forget Tissue-culture that-'Just' KING-Works!!! >(*U^)<

Although It Hasn't-Been Much-of R 'Work'-Day, so-to-speak ... What-with the DEATH-of Osama-Bin-LADEN ... "I"-Did Managed to Get-down to The-Shops Sometime IN-The-Afternoon &-DECIDED to-Go with-'Glass' Initially Since-It Can-Be ACID-'Washed' if-Necessary ... & the-Lids had somewhat-of-R-"Seal' ... though How-Long Under Repeated-Use They'll-'Last' "I" don't-'Know'!!!??? >(*~*)<



They-R 'Dedicated' [Thin-Walled] Preserving-Jars at-About $2.50-Bucks Fifty R-Piece ... so-Presumeably They-R Borosilicate &-'Should' Do-some-form of -Mileage' Overtime so-to-Speak.



'Got'-My Calculator-OUT & Worked-OUT that "I"-Need / Require about 27grams of-Sucrose [Sugar] per 600mls to-Gain an Osmolarity of ~44g/Litre IE TWICE The Potassium-Amended RHIZOSPHERE 'Salt'-INDEX of Plants-like Drosera-binata, Dionaea Etc..



Unfortunately although 2ccs of-Agar Made-UP to 600mls Appears to-Be 'Ideal' for the Work "I"m-Attempting ... My 'First' Error, Mistake or TEETHING-Problem IN Getting-'bach'-INTO The-Swing-of-things Was-to 'Forget' that Such-R-Large Quantity-of Sugar Would 'Rob' Some-of The 'Heat' Away from The 'Pot' / Crucible or cCoking-Vessel ... &-So 'Not' ALL-of The Agar Dissolved this-Time. the-Solution of-course Is-to Either Pre-WARM The-'Pot' &-Or Dissolve-In The Agar FIRST 'Before' Adding The Sugar 7-Then Fully making it-UP to 600mls. >(*~*)< / >(*U^)<

As-Mentioned Before "I"-Want-R SMACKBANG / Set-&-Forget Method of TYNDALLIZATION That's Time-Poor & Savant-Heavy so-to-Speak If-At-All 'Possible'. >(*U^)<



Anyway "I"-Whacked-UP Two 'Inital' Qualatative Lines 'Just' to Test-the-Waters so-to-Speak.



'Did' the GLADWRAP-Thing to-'Supposedly' Lessen or-Negate Airflow-&-Mites.



One-Jar of-Each "I"ll Leave, as-IS ... to-Determine Rhough 'Contamination'-Time.





The-Other "I"ll 'Begin' TYNDALLIZATION-On Tomorrow to-determine the Intial pitfalls &-Whether-or-'Not' The-Process has-Any Merit Within The-Rhelms of Tissue-Culture per-se.





Anyway "I"ve Placed ALL-Jars IN-R-'Top' Ceiling-Cupboard for Regular-'Warmth' to-'Hopefully' ENCOURAGE Spore-Germination ... The-'First' Crop-of-Which "I" Will-Attempt to-Kill Tomorrow with-Our 'First' TYNDALLIZATION IN The Household Laundry-Trough Water-Bath so-to-Speak!!! >(*U^)<

It'll-be 'Int'resting' to-See Whether-or-'Not' The Agar Remelts at-Around 60oC.

FINDING R Small 'Cheap' Set-of Thermometers "I"-Could Place Within The Jars to-Determine Whether-or-'Not' "I"-Reach The Critical 53oC INSIDE The-Jars Has-Been 'Fruitless' so-Far.

PERHAPS-Tomorrow??? >(*~*)<

Yeah ... 'Most' Laundry-Troughs Arn't-PRETTY ... Since-They're Basically Only 'One'-Pipe-AWAY from-R-SEWER so-to-Speak. >(*U^)<



Anyway ... 'Ran' The-'HOT'-Tap for-about 35-Seconds Until The-Stream Was:



SCALDING IE ~60oC. >(*~*)<



Then 'Stopped' The-Plughole, Placed Two-Jars IN-The-Trough 'Ready'-for Their 'First'-TYNDALLIZATION.



Place-R Low-IQ Housebrick ON-Top for Good-Measure &-'Protection' from Possible Unwarrented Boyancy Negotiations.



&-Then Proceeded to-Fill The-Trough With 'Hot' SCOULDING-Water UPTO The Level-of the GLADWRAP On-The-Jars so-to-Speak.



The-Water Was Now too-'Hot' to-Keep-Ones Hand IN-The-Water ... &-'Surprisingly' Remained Quite-'Hot' for-at-Least Half-an-Hour ... Which "I"-Hadn't Anticipated and-Didn't Come-down to 'Ambient'-Temperatures for Well-over an Hour-&-A-Half At Which-Time "I"-'Discovered:



The-'Brick' Per-se Was Totally-Unnecessary With-such 'Thin'-Walled Preserving-Flasks ... as-Well-as the-Fact that The-Agar Hadn't 'Melted' so-to-Speak ... Which-IS R-Plus IN 'My'-Book!!! >(*U^)<

****
Anyway My-Idea IS-to keep TYNDALLIZING IN-'This'-Manner ON-R-Daily-Basis On The 24-Hour-Mark UNTIL One-of The 'Control'-Flasks 'Fluffs'-with Fungal-Hyphae ... We-Will-'Then' Know Whether-or-'Not' Sulphur has-Made Any-IMPACT On 'Delaying' Spore-Germination / Hyphal-Development as-Well-as Whether-or-'Not' TYNDALLIZATION Has-Made Any-Impact On-Either-Line so-to-Speak ... from Which Information We-Can Design Future Experimentation.

Oh... okay?
I'm just gonna pretend I know what all those posts where about!

Well-'Dave' ... TODAY "I"-'Started' Playing-around with The Major-MACROS (Macronutrients).

'Hopefully' These-will Help The 'Control'-Flasks to-'Fluff' Much-Faster & More-Reliably ... for until They-'Do' "I"ve-'Got' NO Yard-Stick, so-to-speak ... to-Measure-anything Against Other-than 'Experience'. >(*~*)< R-Bit-like Saying You've 'Killed' The-Worlds Most-Wanted &-'Not'-Being Able to-Produce R-Photograph!!! >(*~*)< / >(*U^)<

'Until' One-of The Control-Flasks 'Fluffs' ... "I"m Encumbered-by This Nagging-Feeling that Shaking-UP The Medium IN-The-Jars While 'Still' Warm May-have Inadvertantly Despored The Vessels. One-Is Not 'Suppossed' to-Do THIS since-it Repudedly Leaves Sugary-Medium In-The Screw-Threads of the-Jars Through-which Fungal-Hyphae can-Grow ... but Then-again TYNDALLIZATION 'Should' Kill Any-such Hyphae ... &-Of-Course EXTINCTION Is-'Forever' ... so-R NEW-Spore Would-have to-Germinate for new-Hyphae to-Eventuate.

HOPEFULLY ... 'Tyndallization' ON-R-Daily-Basis 'Around' The 53oC-Mark May-be The-Solution We've ALL-been 'Looking'-for: Little-Damage to Large Explant-Material ... but With R-Continual Daily Clean-UP or-CHECK on Burgeoning Bacterial-&-Fungal-Infection. The-Explants Continually Has 24-hour Periods to (Re)-Build NEW Cellular Material for Damaged-Cells ...but the Bacteria-&-Hyphae R-DEAD so-to-speak!!!??? >(*~*)< / >(*U^)<

Having ALL Your Flasks IN-R Large Water-Bath that-You Fill-UP Once-Daily May-be The Daily-Grind ... but it-Could-be Easily 'Automated' If-'Successful'.

But-'Thart's Probably Putting The-Cart before The-Horse as-Yet. >(*~*)< Let's-'Just' SEE-how We-Go
FIRST With the-'Basics'!!!>(*U^)<

Well ... The Results R-IN from Stage-I Trial



"I"ve-'Tried' CANDL'E'LING The-Jars for-Your Erudation ... but-"I" 'Think' "I"-Need to Perfect The-Set-UP Before-"I" End-UP Posting any 'Useful'-Images Your-End so-to-Speak.



OK Here-Goes:

(1) No 'Visible'-FLUFFING Above Surface-of-Media Even-After 10-Days ON-ALL-Jars ... Including-CONTROLS.
(2) ALL-Contamination 'Appears' to-Become Evident IE MACROSCOPIC ONLY-After at-Least 7-Days
(3) MACRONUTRIENT-Additions Appear to-INCREASE 'Vigour'-of-Contaminations.
(4) TYNDALLIZATION Does-Appear to-Have A-COUNTER-Effect ON-Contamination as-Well-as Their Subsequent 'Vigour'.
(5) SULPHUR Does Appear to-have a COUNTER-Effect On-Contamination but Not-Necessarily 'Vigour' Per-se.
(6) TYDALLIZATION IN-Conjuction with SULPHUR Appears to Limit Contamination (So-Far) After 10-days to just One-Organism IE to-What Appears to-Be R-'Grey' Penicillium-species.



So-Far 'Grey'-Penicillium Colonies R-Limited to-'Just' R-Managable 3-5 Colonies ... Though Doubleling The-Concentration of-Sulphur or Adding some-Other Retardant May Completely 'Sterilize' The-Jars / or-Effectively Limit-Growth of ALL-Organisms When-Combined with 'Regular'-TYNDALLIZATION???



The-'Use'-of-The-Term REGULAR Needs to-be Defined ... Because it-Was 'Found' After 'Missing' R TYNDALLIZATION Due-to:"Mothers'-Day" ... that Apparently 'Missing'-R-Day Was-'Not' very-Detrimental to The-Overal Skeme-of-Things so-to-Speak ... &-That TYNDALLIZATION Ought-to-Be Primarily 'Thought'-of as-R Management-Tool Rather-than an-Overriding, ALL-Conquering Procedure Per-se.



Anyway:... WHERE-to-Go From-HERE.

The-Next-Stage Would-be to EXPLORE The-'Effect' of Regular-TYNDALLIZATION On Large-Plant Explant Material so-to-Speak ... & Whether-or-'Not' Large-Pieces-of-Material R-Able to 'Cope'-with as-Well-as 'Recover' Dead-Cell Material Within The Regular 24-hour Framwork of the Procedure Overall &-Whether-or-'Not' We-Can 'Trick' Plant-Material Into-'Believing' They-R Within The-Confines of-R Tropical SAHARA-Desert Type/ Like Arrangement WITNIN-The-Jars, Themselves, so-to-Speak With-'Seemingly' Long CHILLY-Nights Regularly-Followed by Short Steamy ROASTING Days!!! >(*~*)< / >(*U^)<

This is the most bizarre post i have ever seen. Richard please message me and i will at least give you a basic outline of the tissue culture method.