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Offlineoxana
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Registered: 12/14/08
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Re: Culturing • GRAIN 'PETRI' TEK [Re: WillSolvem]
    #19284418 - 12/16/13 03:08 PM (10 years, 1 month ago)

Quote:

WillSolvem said:
To stop condensation in either agar or grain.

First condensation occurs only in the cool side of things where they're is moisture already present. So if your media (agar, grains, ect) has a temp around 75°f but an air temp of 70°f you will have condensation. To remedy try increasing the air temp around the media, remember that with increased temp you will have increased evaporation

Second properly hydrated media is key, take agar for example, after you pour hot agar it continues to out-gas for several hours even after cool (don't believe me, just sit a hot agar dish on a scale and record it's weight over a few hours). To remedy this I (personally) will mix, heat, and pre-pour my agar into dishes, let cool and to firm up, then sterilize. Let the autoclave completely cool before opening. With grains I make sure they are room temperature before putting lids on and sterilizing.

Hope this helps :thumbup:




thanks ill try that


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OfflineBigGreenMat
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Re: Culturing • GRAIN 'PETRI' TEK [Re: oxana]
    #19300510 - 12/19/13 11:12 PM (10 years, 1 month ago)

Hey Violet, I have a question for you about your in vitro tek.  I have 2 of the Quart containers fully colonized for almost a week now.  How do you sterilely bottom water?  I have some autoclaved tap water.  Should I just try to do it sterily my best in an SAB or should they pin and fruit without watering at first so I can get my first pins out?

Also as an update.  I have VERY little growth from the Pan Cyan.  Seems to not like straight brown rice... I am considering taking one of the colonized grains and moving to an agar plate to see if that will move things along.  In another thread I was told that Pan Cyan should pretty easily colonize grain and only needs to be cased or spawned with poo.  Maybe I just have some bad spores/genetics.


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InvisibleViolet
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Re: Culturing • GRAIN 'PETRI' TEK [Re: BigGreenMat]
    #19303465 - 12/20/13 03:24 PM (10 years, 1 month ago)

Don't water until after the 1st flush.  The cakes may not have enough water for a full-size 1st flush, particularly the extra-shallow cake dishes, but that's kindof the point of the tek as well.

You don't have to water them sterile.  It could only help to do so if you intend to continue having totally clean clones or prints to take after the 1st flush.  Otherwise it's nearly pointless to use sterile water to water as clean procedure!



"In another thread I was told that Pan Cyan should pretty easily colonize grain and only needs to be cased or spawned with poo."

It's interesting how some pieces of information hold to be both very true and quite untrue depending on the context and how the statement is taken.
Nobody can convince me that the different grains are just the same to mycelium, especially to the more picky mycelium.  All these species can grow on grains not because all of them are natural substrates but because of similarities to their natural substrates.  Mycelium can colonize brown rice and rye seeds because of their similarities to the grains of other grasses.
It makes good sense to me that you could have better luck with Panaeolus using grass seed than brown rice.  You should be able to get Pan mycelium to grow over anything once up and rolling... but perhaps you'll find better culture luck starting with new spores on grass seed, grass seed agar, or dirt agar (+gs water) as germ media.
Combined with that, I feel that it's moreso the Casing that helps Paneolus, less so a substrate "requirement".  If grass seed and casing fulfills the needs, then the "benefit" of using poo is overrated.  I've definitely seen many great Pan grows that use very little poo in the sub mixture.


--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
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InvisibleViolet
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Re: Culturing • GRAIN 'PETRI' TEK [Re: Violet]
    #19303500 - 12/20/13 03:29 PM (10 years, 1 month ago)

On a side note  (pun intended)....  I'm starting to get pretty fed-up with straight grains invitro.
They don't easily seem to pin where I want them to on uncased grain cakes, and although this has been a tiny bother it has been extremely educational.
I've learned that Cubensis does not want to fruit from highly-nutritious spots if it can avoid it!

So many of the pins from my single invitro culture tests formed up on the side of the plastic container instead of from the rice cakes themselves, and when they formed on the cake it was usually on the side/bottom in the "microclimate" despite the invitro container retaining humidity well so the whole container is a good climate.
It was confusing seeing only the extremes of "courage" and "cowardice" both coming from every one of my many isolated cultures... but my experiments with Anne Halonium's pom tek shed some special light on this for me.  This behavior is instead the ideal fruiting behavior when a low- or no-nutrition pinning surface area is provided.


I believe this offers a greater perspective on side-pinning.

Pinning seems much more likely and numerous at "barriers", regardless of even if the mycelium just got there!  Mycelium doesn't necessarily want to fruit from where it first started colonizing if it's not a great place for it.  Almost every one of my clones from "grain petris" were taken from the most recently colonized edge of the shallow cakes, from mycelium that was only a week-ish old!

It's well known that "the #1 pinning trigger is full colonization of substrate."  But I think this is not exactly as it reads, but rather we think of it this way almost entirely just because of how we come at cultivation, since we put the mycelium in a "box" of conditions so-to-speak.
Some curve balls in cultivation show us that it's not quite as said:

They're pinning exclusively at the edge of where they have not colonized!

Also, all of my "pom" grows pin Far more from the scrubbies which are expended of nutrition shorty after colonized,  fed by migrating water and nutrition from the nutrient-rich grainwater slurry below.

This is why I think grain cakes benefit so hugely from a casing layer.  It's a sort of low-nutrition "barrier" that allows the mycelium to fruit away from their nutritive source.  It's not JUST for moisture retainance, and not JUST for creating an ideal microclimate, but about giving Cubensis a low-nutrition site on which it will love to pin, instead of forcing it to pick the best of the worst fruit sites on the sides of highly-nutritious cakes.

And it sure matches my experience.  Using a casing layer seems almost like a magic trick for getting Cubensis to fruit fully from straight-grains in fruiting conditions.  Otherwise they seem much more likely to "do nothing", despite all the potential of the highly-nutritious substrate they're on!

Don't anybody ever tell me again that Cubensis doesn't "need" or even really "benefit" from a casing layer.  It does.
Such a difference may not so visibly manifest with "bulk subs" because they're much more nutrient dilute, but this is not an "advantage" to bulk substrates but another example of how the use of bulk substrates hazes myco perceptibility.

A casing layer, or other provided nutritionally-dilute fruiting surface, has Huge benefits for Cubensis particularly and especially on substrates packing lots of nutritive potential.  It directs what we have thought is the desire to "side-pin" towards our intended fruiting surface, and seems to 'unlock' the full fruiting potential of all that dense nutrition, whereas otherwise results may seem pathetic and extra-slow while also annoyingly coming mostly from the sides since the mycelium cannot find a location to fruit that best suits their preference.


Large yield is made possible not only with lots of nutrition but by providing an ideal fruiting surface away from the nutrition source for that nutrition to be used.  A cake with large nutrition can still easily have shitty yield if the two are not brought together.


So, to bring this full-circle...  I suggest using casing layers even growing full half-pint cakes invitro, with extra-loose lid!


--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
Violet's Teks and Posts


Edited by Violet (12/20/13 03:38 PM)


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Invisibleanne halonium
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Re: Culturing • GRAIN 'PETRI' TEK [Re: Violet]
    #19303640 - 12/20/13 04:00 PM (10 years, 1 month ago)

hmmm.......
some of that stuff has me perplexed violet.

normally i see very few side pins.
usually i case. but not always.



even when i dont..........

keep in mind, LED lighting is highly directional
and these are all LED pinsets.


--------------------
:aliendance:


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InvisiblePastywhyteMDiscord
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Re: Culturing • GRAIN 'PETRI' TEK [Re: anne halonium]
    #19304132 - 12/20/13 05:48 PM (10 years, 1 month ago)

From what i have seen cubensis and pans tend to pin from the sides of the substrates in nature and on a sub indoors you can only get a good top pinset if you deny all other avenues for pin formation. If you allow side pins to get a foothold it seems that the culture will pour all its energy into those spots to the detriment of other seemingly viable options regardless of the conditions.

If you ever see people fruiting bulk cakes or brf cakes, most of the pins form from the sides despite te fact that the verm or casing application is usually better on top. I do have some experiments planned down the road to test this further.


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InvisibleViolet
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Re: Culturing • GRAIN 'PETRI' TEK [Re: anne halonium]
    #19308646 - 12/21/13 04:33 PM (10 years, 1 month ago)

That's great Anne but it hasn't worked out for me like that, even with the cultures that have turned out to be best so far.


I didn't get (m)any photos of the first pinset which had many pins from the rhizo networks on the plastic, but this is in effect what I almost always get:


None on the bare surface of the grain itself.

Even in the seemingly best of invitro conditions.


More pins from this wedge stuck on the side of the plastic than from the rest of the cake put together:

I find that particularly telling about fruiting preferences, nutritional dilution, and agar growing ;]

It's not that I'm not getting full yields and efficiency after all flushes...  Because I am, still the best I've seen even multi-spore.

It's just that those yields come almost exclusively from the edges of the cakes.  Not really a problem with the smaller invitros like 'grain petris', but a pain-in-the-ass for full sized ones, and in some instances the reduced total fruiting has effected yield.


Yet the same behavior on Poms is actually a Great thing and is one of the biggest advantages of Poms I've seen yet.

I'm becoming a full-on Pom convert.  Damn it works so well.


--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
Violet's Teks and Posts


Edited by Violet (12/21/13 05:25 PM)


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Invisibleanne halonium
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Re: Culturing • GRAIN 'PETRI' TEK [Re: Violet]
    #19308771 - 12/21/13 05:04 PM (10 years, 1 month ago)

shhhh.
poms, youll spark WW3........:thumbup:

i am sorta baffled at the side pin thing though.


--------------------
:aliendance:


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InvisibleWillSolvem
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Re: Culturing • GRAIN 'PETRI' TEK [Re: anne halonium]
    #19315733 - 12/23/13 11:15 AM (10 years, 1 month ago)

Pins!! I'll be back in a few with the porn :thumbup:


--------------------


AMU Q&A Thread because questions deserve answers.


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Invisibleanne halonium
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Registered: 05/07/13
Posts: 1,908
Re: Culturing • GRAIN 'PETRI' TEK [Re: WillSolvem]
    #19316095 - 12/23/13 01:10 PM (10 years, 1 month ago)

Quote:

WillSolvem said:
Pins!! I'll be back in a few with the porn :thumbup:




rockin........:thumbup:


--------------------
:aliendance:


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Invisibleblindingleaf
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Re: Culturing • GRAIN 'PETRI' TEK [Re: anne halonium]
    #19317694 - 12/23/13 06:37 PM (10 years, 1 month ago)

so i have a couple violet tek things going
(two middle containers will be addressed below, as for the coffee filters on jars, they are a dust buffer, there are SFD's hiding underneath, just in case someone was about to chime in with a "YOU USE COFFEE FILTERS!?!?!  i have 2 cats and a dog, plus i smoke in adjacent room, so a lot of dust is generated)



some are "grain petris" and 3 are filled with whole BR, and plan was to use as spawn.  so i guess i used violets PP5 method and her BR prep method in the case of these 3 containers.  problem with these was I over saturated the BR.  innoc'd with agar wedge, and growth was going, just slow because of excess moisture. 



so just now, i could not help myself and shook 2 of the three containers that look like the one above. i know violet tek is not a shake needed tek, but i have many grain jars going, so if it didn't work I have more than enough to keep me busy. my goal was to spread colonized grains around and increase colonization speed as well as maybe better distribute the excess moisture around grains and walls of PP5.  but the most curious thing happened, 1 min after i shook i looked up and saw blue!!!  never has this happened to me



a chill went down my spine at first because i though there was a contam hiding on inside and the shake dispersed it, but its just super blue now....anyone else had this happen?  is it because the grain is so soft that it is easy to bruise?


--------------------
A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!

The whole is greater than the sum of its parts


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Invisibleanne halonium
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Re: Culturing • GRAIN 'PETRI' TEK [Re: blindingleaf]
    #19317736 - 12/23/13 06:45 PM (10 years, 1 month ago)

they will do that sometimes.
especially if ya used ferts.

the big thing with shaking PP5, is that because they are "soft"
shaking changes air pressure in them, and if your not under best conditions......bacteria.

as far as water? sterile water is harmless.
the poms prove your not gonna drown your mycelia,
and the airflow/ FAE is more forgiving in PP5 .

therefore, water is not an issue.
if ya cooked em right in PC, and had a clean innoc.
water will affect nothing.

over all, looks good though.


--------------------
:aliendance:


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Invisibleblindingleaf
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Re: Culturing • GRAIN 'PETRI' TEK [Re: anne halonium]
    #19318073 - 12/23/13 07:48 PM (10 years, 1 month ago)

hmmm...well like my post said, the jars and PP5's share a room with a dog and 2 cats...hopefully the shake will not breed bacteria if it did indeed change the air pressure.  if it did its ok, i have one more BR that was not shaken, as well as several that are only 1 inch high, like the grain petri tek.  i actually just nocced up three more with an lc started with agar.  anyone have a pic of an agar petri with myc AND bacteria?  i've been having some bacteria problems in my regular glass quarts, even with SFD's, and its either because my WBS was not PC'd long enough or there was an unseen bacteria on my plate.  my sterile technique is fine because all of my agar work is clean (unless there is some very translucent bacteria I'm not seeing, thats why i think its the PC time for WBS-just 60 min). another thing that makes me think its PC time is the 3 jars that did get bacteria colonized to 80% and then the last 20 had no growth and the tell tale slimy/wet grains against the jar wall. just switched to rye though, i like it better just cause i'm bad at getting WBS moisture correct even with 24 hr soak/10 min simmer. they are either too dry or burst.  I've done it with success before, all my grows have used WBS, but now that I'm starting more and more jars, i didn't want to have 1/10 fall to bacteria.  bad side is price of rye...luckily my friend works at whole foods, so the 20% discount helps, but still not close to price per lb of WBS.


--------------------
A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!

The whole is greater than the sum of its parts


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InvisibleJ. Jack Flash
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Registered: 11/20/13
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Re: Culturing • GRAIN 'PETRI' TEK [Re: blindingleaf]
    #19318621 - 12/23/13 09:36 PM (10 years, 1 month ago)

thought i'd throw some pics of my work on this thread, for giggles.


my first MS inoculations on agar, transferred to grain petri a week ago.  kept the jars to see if they'd contaminate, as i felt like i really bungled the process.  look good to me so far.


next are the grain jars to which the samples were transferred.  moving right along.  the last two are the same jar.  they look so scattered because that's the jar i had problem with.  couldn't get the wedge to let go of the salpel, flung it across the jar, stuck to the side, had to dig and dig and dig to pick it up and put it in the middle.  such a noob.  :rolleyes:


finally, MS direct to rice made at the same time as the agar above.  watching these progress has been absolutely beautiful.  i'm loving this mycology thing.  three jars originally, one lost to trich i think.  tiny green dot, out it went.


--------------------
 
the j stands for jesus.
2020 new years grow along


Edited by J. Jack Flash (12/23/13 10:07 PM)


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Invisibleblindingleaf
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Re: Culturing • GRAIN 'PETRI' TEK [Re: J. Jack Flash]
    #19319835 - 12/24/13 06:35 AM (10 years, 1 month ago)

awesome JJ!  haha i still get the agar stuck to the scalpel, its annoying!


--------------------
A few thoughts on cultivation
MICROBIAL HUSBANDRY!!!!

The whole is greater than the sum of its parts


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Invisibleanne halonium
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Re: Culturing • GRAIN 'PETRI' TEK [Re: J. Jack Flash]
    #19319846 - 12/24/13 06:41 AM (10 years, 1 month ago)

ok JJ looks good to me.............


--------------------
:aliendance:


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InvisibleWillSolvem
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Re: Culturing • GRAIN 'PETRI' TEK [Re: J. Jack Flash]
    #19320443 - 12/24/13 10:28 AM (10 years, 1 month ago)

Quote:

J. Jack Flash said:
couldn't get the wedge to let go of the salpel, flung it across the jar, stuck to the side, had to dig and dig and dig to pick it up and put it in the middle.  such a noob.  





You will find at a pet supply store those reptile heating pads, buy the smallest one and tape to the underside of your SAB. When setting up for agar work plug it in, set up your SAB but place your scalpel over the area with the heat pad underneath, by the time it's closed, sanitised, and got your gloves through you have a nice warm blade for agar to slide right off.

Like a warm knife through butter :wink: :thumbup:


--------------------


AMU Q&A Thread because questions deserve answers.


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InvisibleWillSolvem
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Re: Culturing • GRAIN 'PETRI' TEK [Re: WillSolvem]
    #19321975 - 12/24/13 04:10 PM (10 years, 1 month ago)

Two of the containers started pinning so I decided to take clones

This one I'm calling Santa



And this one Fatboy





Sorry I had compress the rest of the photos, getting close to my weekly upload limit

Santa




Fatboy




Fatboy was actually sooo big I had to stab it with the tweezers to remove it! :thumbup:

I will keep you all posted while I grow out these two strains

Merry Christmas Everybody :cheers:


--------------------


AMU Q&A Thread because questions deserve answers.


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Offlineoxana
i am the fun guy

Registered: 12/14/08
Posts: 258
Last seen: 2 years, 14 days
Re: Culturing • GRAIN 'PETRI' TEK [Re: WillSolvem]
    #19324062 - 12/25/13 01:32 AM (10 years, 1 month ago)

so im looking to isolate and test out some new strains to be used in monos.

im wondering if i fruit an isolated strain on several brown rice pucks and they do well on brown rice, will this be a pretty good indicator that this strain will also do well on coir/verm/grass seed?

it seems like a much easier path to take then to waste limited monotub space on strains that are not proven to provide high yields and other positive factors.


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InvisibleJ. Jack Flash
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Registered: 11/20/13
Posts: 1,500
Re: Culturing • GRAIN 'PETRI' TEK [Re: oxana]
    #19324333 - 12/25/13 03:45 AM (10 years, 1 month ago)

i'm not experienced enough to answer the question definitively, but can say for certain that this is what i'm counting on.  also, it's pretty much my understanding of the whole point of this approach... that is, to draw out and test isolated Strains using as little time and as few resources as possible. 

thus achieved, you're ready to improve the world with whatever larger scale cultivation method you desire.

:growshrooms:


--------------------
 
the j stands for jesus.
2020 new years grow along


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