soma_seeker

Hi guys, Chasing a clean cubensis print for microscopy purposes, ideally for the weekend. Happy to pay for item and express postage Cheers

Hi all Found myself in a bind and need a fresh cube print for the weekend for microscopy purposes. Will happily reimburse nicely anyone who could express mail me a recently acquired print tomorrow for overnight to SE QLD so that I'll have it for the w'end. Cheers

Sorted Thanks.

An express envelope should reach SE QLD from most areas Friday if sent tomorrow.. Cheers anyway though

Hi everyone, I just wanted to share my design for a low-cost, small laminar flow hood. This hood was designed to aid in simple grain to grain transfer and agar work. I built this unit only several weeks ago, but I can report that after numerous attempts at both of these types of work, I have a ZERO contamination rate Prior to building this hood I was using a glove-box which was highly cumbersome, impractically small for grain to grain transfer work and still resulted in regular contamination when working with agar. Firstly, much of the credit for this work must go to Endorfinder for first alerting me to the fact that a home-made flow hood could in fact be prepared for less than the usual $400-500 price range. For those of you unfamiliar with the common design of a simple home-made hood, there are three key pieces; a pre-filter, a HEPA filter, and a fan. Endorfinder alerted me to a particular fan, which lowers the cost of the unit considerably but still provides the necessary function. This fan is available on eBay for $75 delivered. Generally, the fan is the most expensive of the three items, followed by the HEPA filter. In my (albeit limited) research, the cheapest HEPA filter I could find was approximately $100. This filter was an inch thick and filtered all material of a diameter greater than 0.5 micron. After scouring the net I then found this filter for $46. The dimensions of this unit (which is what gives you your effective working area), are 36cm by 27.5cm. It is sold as a replacement for a specific household air purifier, it is about an inch thick and is said to filter all material of a diameter greater than 0.3 micron. It should be noted that it is best to use a HEPA filter which is as deep as possible, so as to allow for a constant and steady outflow of air across the filter's surface. Using a one inch thick filter is therefore not ideal, but the goal of this design is to balance function with affordability (and as I have said, it has functioned very well for me so far). Also, the dimensions of this filter give a fairly small effective working area. I have found it to be sufficient, but i would not attempt anything smaller. For my pre-filter I purchased the pre-filter of the same model of air purifier. It cost me $11. I had intended to use this filter as is, but in the end I realised that it was far more appropriate to cut it to a smaller size. That being said, you might be able to find something cheaper that will do the job, maybe even something lying around the house (but this filter does do the job very well). The remaining items you will have to purchase are; 25mm wood screws (approx. $5), a combination glue/sealant (approx. $10) [NOTE: do not buy a simple silicone, buy a quality combination material that acts as both a glue and a sealant like this. Silicone will not do nearly as good a job in this application], 12mm thick plywood for the box (approx. $30 from Bunnings). So, you will require; - Fan $75 - HEPA filter $46 - Pre-filter $11 - Timber $30 - Glue/silicon $10 - Wood screws $5 Once you have obtained these items it is simply a matter of cutting your timber to size and assembling the components. For this you will require a power drill and a jigsaw (in order to cut a circular hole for the fan). I made the mistake of cutting my timber to size before my HEPA filter actually arrived. I used the quoted dimensions of the filter, which actually turned out to be a few mm larger than quoted. This resulted in the filter being slightly squashed in the frame (evident from the wavy pattern on the filter's surface). Obviously there is no need for you to make the same mistake. The (correct) dimensions of the pieces required (when using 12mm thick plywood) are; 2 x 30.2cm * 45cm (top and bottom) 2 x 27.6cm * 45cm (vertical sides) 1 x 36cm * 27.6cm (middle divider) 1 x 30.2cm * 38.6cm (back) Once cut to size, drill or jigsaw a hole in one of the sides at the bottom back position. This hole is for the power cord of the fan to exit. Place the cord through this hole and then, after applying a line of the glue/sealant along the join, screw the sides to the base. At this point I checked to verify the middle divider had been adequately cut to size, by placing it in position as shown in the photo below. Once I was confident that it was all flush, I traced around the diameter of the fan on the divider and cut this portion out using the jigsaw. I positioned the fan such that the small protrusion from the cylinder sat flush against the bottom of the box. This helps to hold the fan in position later. The following photo shows the positioning of the filter, divider and fan. Note that each of these three items have not yet been permanently fixed in position. The order this is performed in is important in order to properly seal the hood. The fan, divider and filter were then removed from position and, after applying a large line of glue/sealant along each edge of the HEPA filter, it was returned to its position (5mm back from flush with the edge of the plywood). Glue/sealant was then applied to the top of the vertical sides of the box and the lid was then screwed into place (optionally, you can attach a handle to the lid before screwing it into position). At this point it is now important to thoroughly ensure that all edges of the box are airtight. This is performed by applying the glue/sealant to each of the edges on the inside of the box (the outside is done as well, however it is important to do the inside while you still have full access to it). Pay particular attention to the edges between the HEPA filter and the box, if any air is allowed to pass through this area then the entire unit is compromised. Once you are confident that these seams are airtight you can put the divider in place, 17cm back from the end of the box. If you have prepared the box well it will likely be a snug fit and you might have to use a rubber mallet (or something similar) to knock it into place. Once in place you can drop a couple of screws threw the side to secure it, then seal the seams with the glue/sealant (however from now on the seam work is not as vital as it was when working with the HEPA filter compartment). You can now put the fan in place, such that it will rest flush against the back of the box, and apply the glue/sealant between the fan and what is likely a somewhat rough hole cutout. The cylindrical shape of the fan itself isn't perfect, so don't expect to have prepared a perfect hole into which the fan will fit snugly. Simply use a generous amount of the glue/sealant and it will be securely held in place. Now you can attach the pre-filter to the back of the box. To do so cut the pre-filter down to the shape shown in the image below. Then use the jigsaw to cut a rectangular whole in the back piece of the box which is around 1 inch shorter in length and breadth than the pre-filter. Attache the pre-filter using the glue/sealant, then attach the back of the box using a line of glue/sealant and screwing it into place. Now that the hood is fully assembled it is a good idea to go over all the seams of the box in order to seal any air-leaks, whilst focusing on the front portion of the hood. Pay particular attention to the seam between the HEPA filter and the box. The filter was installed 5mm back from the edge of the box, this was to ensure adequate space for a generous portion of glue/sealant, so as to ensure that no air whatsoever can escape from this space. Now that you have finished, allow the glue/sealant to dry overnight before turning the unit on. The next morning setup the hood on a desk in a room with no drafts/airflow and turn it on. Test the outflow of air by holding a lighter about 2 inches in front of the surface of the filter. The flame should bend very noticeably. As you move the flame around the surface of the filter it should remain bent and hopefully not vary much in its position (indicating a sufficient and constant outflow of air from the entire filter's surface). This hood (like all hoods) will work best in a room that is as small as possible. Before starting your work ensure that all surfaces have been sterilized with 60-70% alcohol, you have cleaned the airspace with an aerosol disinfectant such as Glen20, and that you have had the room sealed with the hood running for at least an hour in order to 'scrub' the air. Final note: Please bear in mind that this design is far from a professional laminar flow hood. The design has several key limitations that you should consider; 1. The HEPA filter removes material of diameter greater than 0.3 micron, however the general standard for mycology is a filter that removes anything larger than 0.03 micron. 2. The HEPA filter is quite shallow, which results in a less even and constant outflow of air from the entire filter surface. 3. The HEPA filter is fairly small, providing quite a limited working area. These points aside, this design is a low-cost and functional alternative to larger and significantly more expensive options. From the results I have obtained so far I think it is a highly effective tool for the recreational home mycologist. Ok you're done, go and prep some agar plates or try a grain to grain transfer

Yeah I did mate Hasn't gotten back to me yet. I got one off him previously that I was very happy with. Cheers

Hi people So, through a series of foolish acts I've found myself without a viable cubensis print for my microscopy work... If anyone could help me out I'd greatly appreciate it. I've sent them out for free in the past but am happy to pay as I unfortunately have nothing to trade. I would be particularly interested if someone has a recently prepared PESA print they are willing to sell! :D Cheers

* PRINT NOW AVAILABLE FOR SALE/TRADE. PM ME * Hi all So I know this might be a long-shot, but I was wondering if anyone in the community might have a Psilocybe Galindoi (ATL7) print that they would be willing to sell me for my microscopy studies? I had one in the past, but after not needing it for the past 3 years it seems to have disappeared... Cheers!

After not being able to find a print of this species I purchased one. If anyone is interested in purchasing half of this print (large, aseptic and for microscopy purposes only), please contact me

Hey people So I'm going to start working with horse poo for the first time. What I'd really like is to find an electric turkey roaster to use for the pasteurisation of my substrate! (like this one http://www.ebay.com.au/itm/BELLA-13425-Turkey-Roaster-Oven-18-Quart-Stainless-Steel-/112055999234?hash=item1a170edb02:g:dngAAOSwzLlXhwa9 ) The problem is I can't find them for sale ANYWHERE in Australia, and buying from the US doubles the price due to shipping Has anyone looked into buying one of these or something similar before? Are there any significant alternatives I can look into? I've used simpler hot water/bucket methods before when working with coir, but with poo I'd really like to be more precise... Cheers!

Nice store. Will give him some business I think

Hey guys Just wondering where you buy your spawn bags locally? They can get pretty expensive so I thought I'd try and find the best price. I don't need an injection port. Cheers

Oven bags in the oven, pot on the stove, or in an esky with heated water?

Righto, cheers The oven method is looking good to me at the moment...

Thanks I hadn't heard of that approach. Would be much safer for me too since I have a gas oven (would avoid the risk of putting flammable oven/spawn bags in an oven that has a naked flame). I also have a digital thermometer with a wired probe. Whatever approach I go for I was planning on having the probe in the centre of one of the containers of material. That way I can start my timer when the temp. reads 65 C. What do you think?

Nice. A good friend of mine did his PhD at Penn State. I actually just spent the past year of mine in the US too. I'd thought that the oven might be a viable option, but then I thought that your standard kitchen oven might struggle to maintain a constant and precise temp. within that low a range... A lab oven however

Cheers This is shameful as an Aussie but I don't actually own an esky! (been living abroad the last couple of years) Think this would suffice? I'll be prep'ing substrate for 1 50L monotub at a time. http://www.kmart.com.au/product/table-top-cooler/819333

Fair enough, scrap that idea I guess. It did just seem quite effective and easy, figured it would be worth it in the long run if I could pick one up for $100 or less... Which of these methods would the community recommend? 1. Heating water to temp and transferring to an esky 2. Heating water and sealed bags in a large pot over a gas stove with thermometer and close observation/flame adjustment Cheers

Yeah that was my back-up plan They just tend to be annoyingly weak and problematic if you want to poke a hole in for a thermometer when pasturising... Thanks though!

That's where I just came back from living Thanks for the offer mate but I should be sorted now.

Hi there So I recently moved back to Australia and have found myself in the need P. cubensis spore prints (for microscopy purposes), and would be tremendously grateful if anyone could help me out! I don't really have anything to trade at the moment unfortunately.... but I've donated plenty of prints to others in the past and promise to pass on the favour in the future if anyone is willing to show me some generosity Thanks a lot guys! Peace

Sorry guys, haven't been active for quite a while... In response to your question, yes the unit has been quite effective in maintaining a low contamination rate for the majority of my work. Let me explain, I've continued to have low rates of contamination from; 1. All agar work. Germinating spores, strain selection, agar wedge to LC etc. 2. Syringe prep 3. Spawning colonised qrt jars to more qrt jars (I generally turn one into 8) The one area that I have recently had more issues with however is spawning colonised qrt jars to myc bags of sterile grain. Considering that the level of air exposure (and hence likelihood for contamination) is significantly higher when working with bags than jars I guess I shouldn't be suprised that the contam rate here is higher. However, initially I didn't seem to have a problem so maybe my technique has laxed (probably likely after my initial successes), or I just had a bad recent run. Regardless, I'm still happy with setup and don't feel the need to upgrade/change. If you don't play around with myc bags then my recent problems aren't even a potential issue for you, if you do, chances are you'll have better luck than my recent work Best of luck!

Awesome! Feel free to contact me if you want to verify anything. If not, make sure you let us know how it goes

Still keen to see some photos of your flow box endorfinder Just wanted to report that this design has proved very useful for mycobags. I use the flow to fill the bags with a full plume of sterile air, after which I inoc them via g2g, agar wedge or LC. I then seal them using an impulse sealer in the sterile air-flow. After preparing over 30 bags in this way I've had 2 or 3 contaminate with trichoderma. I'd obviously prefer 100% success, but I'm happy with 90% or better. Now that I have a reliable method for preparing mycobags (something which has always given me trouble in the past), I've all but given up preparing quart jars. My 23qt presto PC can hold 10 quart jars, or 4 mycobags with 3.5L of grain in each. This means that for each PC run I can prepare more than twice as much grain using mycobags rather than jars (based on the quart jars actually containing 650mL each).

No problem mate, no hard feelings here. Nice roses