2015.11.13 Community Chat

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On Dot Plot Exclusion Lab Analysis, Understanding Ensemble Diversity, Partition Function, Positional Entropy, & Vienna Server Prediction Tools

 

DevChat T-05m 

DevChat T-01m 

DevChat T-30s 

Nando: I guess I really should check ChatBot's clock some day...  [2:53 PM]

DevChat T-10s 

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KA-BOOOM!!! 

Elves: :D  [2:55 PM]

Nando: hi elves  [2:55 PM]

Elves: hi Nando!  [2:55 PM]

Jennifer Pearl: hi [2:57 PM]

Elves: Nando my image of you fixing chatbot is now going to be of you peering through a little magnifying glass at mechanical gears, deeply pondering the clockwork [2:57 PM]

Jennifer Pearl: lol [2:57 PM]

Elves: hi Jen!! :D [2:57 PM]

Nando: I foresee myself with a hammer, mostly :D [2:58 PM]

Elves: ahaha can it be a tiny hammer that you hold with tweezers? :D [2:58 PM]

Elves: pling pling pling [2:58 PM]

Nando: brute-force is sometimes a very effective method :) [2:58 PM]

Jennifer Pearl: yes inndeed [2:59 PM]

Jennifer Pearl: hi Elves [2:59 PM]

Jennifer Pearl: !! [2:59 PM]

Elves: hehe [3:00 PM]

Elves: so to answer the previous question, i think that today is a community chat, but if there are any dev matters of course the floor is always open [3:01 PM]

Elves: last week, the monthly das lab presentation was consolidated into the time slot, so we did not have a traditional dev meeting per se [3:01 PM]

Elves: that being said, if anyone has any issues to report we can do that now [3:02 PM]

Elves: like what color is nando's hammer, and does the wavelength of light reflecting from it affect his hammering? [3:02 PM]

Elves: important people want to know... [3:03 PM]

Elves: :D [3:03 PM]

Jennifer Pearl: i have some stuff i have found [3:03 PM]

Jennifer Pearl: in my analysis of the Exlusion and Same state labs including the NG labs [3:03 PM]

Jennifer Pearl: if no one else wants to speak [3:03 PM]

Elves: that would be really cool to hear about!  [3:03 PM]

Jennifer Pearl: dont have alot of my good charts here but I have a couple things [3:04 PM]

Jennifer Pearl: still in the middle of analysis [3:04 PM]

Jennifer Pearl: Sooo that being said I have found that for Exlusion 1-6 and NG 1-3, Same State 1-2 and NG 1-3 if a design has a ensemble diversity over 15 according to Vienna 2 then the design will socre below or right around 80 [3:05 PM]

Jennifer Pearl: and designs that score between 80-100 have divesitys under or equal to 15 [3:05 PM]

Elves: interesting, so a lower ensemble diversity = higher score in this data? [3:06 PM]

Jennifer Pearl: there are designs that score below 80 that have ensemble diversities less than 15 [3:06 PM]

Elves: is it more accurate to say that a higher ensemble diversity signifies a lower score? [3:07 PM]

Jennifer Pearl: yes [3:07 PM]

Jennifer Pearl: when over 15 [3:08 PM]

Elves: is there a range limit for ensemble diversity? like is 15 in the middle or towards the end of the range? [3:08 PM]

Jennifer Pearl: 15 is middle [3:08 PM]

Jennifer Pearl: they go as high at 30 [3:09 PM]

Elves: And for us lab newbies, what is NG 1-3? [3:09 PM]

Jennifer Pearl: the NG labs 1 though 3 [3:09 PM]

Elves: cool thanks [3:09 PM]

Jennifer Pearl: for exclusion [3:09 PM]

Elves: What does NG stand for? [3:09 PM]

Jennifer Pearl: at 30 it is almost always a 30 or 60 score designs [3:10 PM]

Jennifer Pearl: next generation i thin [3:10 PM]

Jennifer Pearl: think [3:10 PM]

Jennifer Pearl: they are labs without predefined structures [3:10 PM]

Elves: at 30 ensemble diversity the score is 30 or 60? is that the score clustering Rhiju was mentioning? [3:10 PM]

Jennifer Pearl: as brourd puts it they are teh build a bear of eterna [3:10 PM]

Elves: ohhh thanks! [3:10 PM]

Jennifer Pearl: yes [3:10 PM]

Elves: ahaha [3:10 PM]

macclark52: What is ensemble diversity? [3:11 PM]

Elves: hi mac! that's a great question, let's find out :D [3:11 PM]

Jennifer Pearl: or very close to 30 and 60 [3:11 PM]

Jennifer Pearl: it is still a new thing for me so I will quote nando [3:12 PM]

Jennifer Pearl: The diversity of that ensemble is a measure of how differentiated or varied the folds will be. In Vienna, this calculation is pretty simple: if Pij is the probability of the (i,j) pair (as shown in the dot plot), then the ensemble diversity is defined as the sum of all Pij . (1 - Pij) [3:12 PM]

Jennifer Pearl: this is from a GetSat [3:12 PM]

Jennifer Pearl: https://getsatisfaction.com/eternagame/topics/designs-with-low-ensemble-diversity-tend-to-score-higher-than-high-ensemble-diversity-what-is-ensemble-diveristy [3:13 PM]

Elves: so it used to be the case that labs were solving for a predefined structure? And now players can decide their own structure in addition to nucleotide sequence in labs? hence nando's glue tool? [3:13 PM]

Elves: thanks Jen! [3:13 PM]

Jennifer Pearl: also from eterna dictionary  [3:13 PM]

Jennifer Pearl: Ensemble diversity - When RNA folds, often it shifts around various different shape. Both ensemble diversity, defect is a measure to indicate how much time the RNA stays in the actual "target" shape. Definition Jeehyung [3:13 PM]

macclark52: That's the Sum - (1-Pij)?  just want to be sure I'm reading the equation correctly.  [3:14 PM]

Jennifer Pearl: sum of all Pij *(1-Pij) [3:15 PM]

macclark52: Ah *.   Thanks.  [3:15 PM]

Jennifer Pearl: here are a couple plots [3:15 PM]

macclark52: Oh, that was a dot.  I read it as a -.   [3:16 PM]

genetech11: user error [3:17 PM]

Jennifer Pearl: https://dl.dropboxusercontent.com/u/87351147/EnsembleDiversity2nd.PNG [3:16 PM]

Elves: so ensemble diversity is the diversity in structure more than nucleotides, though the sequence could also be diverse? [3:16 PM]

genetech11: yes and no [3:17 PM]

genetech11: jk. i didn't even understand that [3:17 PM]

Jennifer Pearl: https://dl.dropboxusercontent.com/u/87351147/EnsembleDiversity1st.PNG [3:17 PM]

Jennifer Pearl: I think that the diversiyt is more structure related [3:17 PM]

Elves: a molecule of RNA is a sequence of nucleotides, which folds up into a structure. that structure itself may shift and change. [3:18 PM]

Nando: it's only secondary structure related [3:18 PM]

macclark52: So that is a measure of the number of possible configurations?  [3:18 PM]

Elves: so if there also happens to be diversity in nucleotide sequence, that is a correlation not a specific factor of ensemble diversity? [3:18 PM]

Nando: @mac: not really [3:19 PM]

Nando: it's about the balance of their likelihoods [3:19 PM]

macclark52: So that one form MIGHT be more likely than another? [3:20 PM]

Nando: the number of possibilities could still be large, but exactly what you just said [3:19 PM]

Elves: if ensemble diversity is the sum of all probabilities of xyz base pairs to form, then a high ensemble diversity means there are many base pairs that could form? [3:20 PM]

Elves: and therefore a wide diversity of potential structures? [3:20 PM]

Nando: the number of possible structures is a factor [3:20 PM]

Jennifer Pearl: I was corious as to whether the lower diversity was casued by less base pairs and I  have found that tthe higher scoring designs (80-100) tend to have 1 to 3 more pairs on average [3:20 PM]

Nando: which is why shorter strands will have lower diversities, generally [3:21 PM]

Jennifer Pearl: but mroe aroiund roughtly 70% of the time [3:21 PM]

Jennifer Pearl: thanks nando that hleps be understand it more [3:21 PM]

Nando: but this diversity measure is about the likelihoods of some structures compared to all others [3:21 PM]

Elves: so one set of base pairs / structures is preferred as the target shape  [3:23 PM]

macclark52: Pij* (1-Pij) is the P that the bond will form x the P that it won't? [3:23 PM]

Elves: and the ensemble diversity is weighted against that preference? [3:23 PM]

Nando: Pij is the probability of the (i,j) pair in the ensemble [3:23 PM]

Jennifer Pearl: i have to step away for a moment so be right back . Here is a chart for your enjoyment [3:23 PM]

Jennifer Pearl: https://dl.dropboxusercontent.com/u/87351147/look%20at%20number%20of%20pairs.JPG [3:24 PM]

Elves: lol thanks Jen! yay charts! [3:24 PM]

Nando: @elves: not sure what you mean... [3:24 PM]

macclark52: So it also includes what's going on with the neighbors.  How in the world do you measure that? [3:24 PM]

Nando: it's a simple computation based on the partition function [3:25 PM]

Elves: my goal is to understand what low vs. high ensemble diversity means.  [3:25 PM]

Elves: so if it is not just a measure of total probability for many or few structures / base pairs to form, how does the "likelihood of some structures compared to others" factor in? [3:25 PM]

Nando: the NuPACK team uses a similar but different concept called ensemble defect [3:25 PM]

Nando: I mentioned it in the GetSat thread [3:26 PM]

Elves: oh thanks i was wondering about the defect reference [3:26 PM]

Nando: as I said on GetSat, for single state problems, you'd normally want to design sequences with the lowest possible diversity [3:27 PM]

Nando: since you want to avoid misfolds [3:27 PM]

Nando: in switches though, you shouldn't be "too good" in that direction (of low diversity), because you actually want your design to form 2 different structures [3:28 PM]

Elves: because low ensemble diversity means lower total probability of different base pair formations, and a single state puzzle should not wiggle into different shapes [3:28 PM]

Nando: well, at least 2 [3:28 PM]

Nando: correct elves [3:28 PM]

Elves: and high ensemble diversity means higher total probability of different base pair formations, therefore increasing the range of possibility for forming multiple shapes / states / structures / base pair bonding [3:29 PM]

Nando: yep [3:29 PM]

Elves: woohoo! thanks Nando :D [3:29 PM]

Elves: so the only part I'm confused is how the target structure[s] factor into the ensemble diversity calculation - directly? or just taking both into consideration? [3:30 PM]

Nando: 2 structures isn't a very high number of them though, so it is somewhat understandable than Jenn found that low diversity designs fare better [3:30 PM]

Nando: the target structures are not a part of that calculation [3:30 PM]

Elves: okay that is very helpful. it seemed the opposite of what I expected, but i think there's a slope depending on just how wiggly it needs to be [3:30 PM]

Elves: wiggly, being a very important technical term :D [3:31 PM]

Nando: the calculation is based only on the partition function, which derives from the sequence [3:31 PM]

Elves: I am new to partition function :D [3:31 PM]

Elves: was that the probability of base pairs to form? [3:31 PM]

Nando: it's hard to explain, but the answer to your question is no, not really [3:32 PM]

Elves: :D [3:32 PM]

Nando: although, each base pair probability is "linked" to the partition function [3:32 PM]

Elves: i'm going through your reply on the forum post, I found this really helpful in understanding why the word ensemble is used: [3:33 PM]

Elves: "the partition function describes how this ensemble of structures will be distributed when equilibrium is reached" [3:33 PM]

Nando: mathematically [3:33 PM]

Nando: can you picture in your mind the set of all possible secondary structures? [3:34 PM]

Jennifer Pearl: im back [3:35 PM]

Nando: now, imagine that each structure is "weighted" by its free energy [3:35 PM]

Elves: a tightly glued together sphere? :D [3:35 PM]

Nando: that's the ensemble [3:35 PM]

Nando: and the partition function is actually just a number, which could be considered as a normalization factor [3:36 PM]

Nando: but I'm getting too technical I'm afraid [3:36 PM]

Elves: it's a total picture of the possible structures according to the free energy per molecule? [3:36 PM]

Nando: sounds pretty accurate, yes [3:37 PM]

Elves: i was wondering literally what is the function, is it what you said "Pij is the probability of the (i,j) pair (as shown in the dot plot), then the ensemble diversity is defined as the sum of all Pij . (1 - Pij)" [3:37 PM]

Nando: that's the ensemble diversity formula [3:37 PM]

Elves: welcome back Jen! [3:37 PM]

Xnessax: hIII!! [3:37 PM]

Nando: the partition function (and the base pair probabilities) are calculated using something else entirely [3:38 PM]

Elves: thanks for clarifying! [3:38 PM]

Elves: hi Xnessax :) [3:38 PM]

Xnessax glomps Nando and Evles. [3:38 PM]

Nando: hi nessa [3:38 PM]

Xnessax: Aw i mistaked it. [3:38 PM]

Xnessax: Hi Elves and Nando! [3:38 PM]

Elves: lol thanks for the hearty glomping. and welcome to the community chat [3:39 PM]

Jennifer Pearl: I have to go. work is calling [3:39 PM]

Nando: bye Jenn [3:39 PM]

Elves: bye Jen, thanks for sharing!! [3:39 PM]

macclark52: Thanks, Jen [3:40 PM]

Jennifer Pearl: glad to share [3:39 PM]

Elves: so Nando is this related to Pablo's talk on entropy? it sounds like similar conclusions in terms of single states preferring lower flexibility and switches preferring higher [3:41 PM]

Nando: entropy is... complicated :P [3:41 PM]

Elves: :D [3:41 PM]

Xnessax: :D [3:42 PM]

Xnessax: You're welcome, Elvinhe" [3:42 PM]

Nando: I can deal with free energies and enthalpies, but I always have trouble with entropy for some reason [3:42 PM]

Xnessax: I don't know how to spell that as the name of cuteness and little-ness.  [3:42 PM]

Xnessax: Oh its dev eh. [3:42 PM]

Nando: but I'd say that it's probably related, yes [3:43 PM]

Xnessax: probabilities) [3:43 PM]

Elves: :) thanks Xnessax. you have a cool name too. [3:43 PM]

Xnessax: Math... [3:43 PM]

Elves: http://prntscr.com/4awkjj [3:44 PM]

Xnessax: No problem, Elvinhe and Thanks, my real name ,did you know? [3:43 PM]

Nando: Eli could probably tell you about the positional entropy outputted by Vienna engines [3:44 PM]

Elves: Nando I think this is the graph I was thinking of [3:44 PM]

Nando: compared to ensemble diversity, it's a localized version of it [3:44 PM]

Elves: wow Xnessax that is cool!  [3:45 PM]

Nando: in other words, domains within the sequence with a high diversity [3:45 PM]

Elves: thanks Nando, I feel like "positional entropy" is a phrase I should know, but I don't know what it means [3:45 PM]

macclark52: I don't either.   [3:46 PM]

Nando: it's the "noise" of possible/probable pairings for a certain base [3:46 PM]

macclark52: clutter in the dot plot? [3:47 PM]

Nando: well, at least, that's the way Vienna uses the word, if I understood correctly [3:47 PM]

Elves: ohhh so just like it sounds then hehehe [3:47 PM]

Nando: ever used Vienna online servers? [3:47 PM]

Elves: no, it would be great to get a quick intro [3:47 PM]

macclark52: No [3:48 PM]

Nando: just paste any sequence in their servers, and with default options, you should find a section about positional entropy in the middle of the results [3:48 PM]

Nando: http://rna.tbi.univie.ac.at/cgi-bin/RNAfold.cgi [3:48 PM]

Elves: here's the guide Eli and I put together on what we got out of Pablo's presentation on entropy, in the early switch days https://docs.google.com/document/d/1FLnV65mAuqx5wFuZXh8Qc-qpNjqh6YLC8XKQwat0QlY/edit#heading=h.kfz5amamscyf [3:49 PM]

Xnessax: Um didnt you know my name, it is vanessa or did you forget [3:48 PM]

Nando: example: http://rna.tbi.univie.ac.at/cgi-bin/RNAfold.cgi?PAGE=3&ID=2XnWbjDC_z [3:49 PM]

macclark52: Oh, vienna is Uni of? [3:49 PM]

Xnessax: XD [3:48 PM]

Nando: scroll down and look below the mountain plot [3:49 PM]

Elves: thanks Nando! no I have not ever seen this, wow cool!! [3:49 PM]

Xnessax: Eh, i trid to get the link [3:49 PM]

Xnessax: theres not a video in live. [3:49 PM]

Nando: the peaks in that graph are positions with high uncertainty [3:50 PM]

Xnessax: From eterna Dev. [3:49 PM]

Elves: Xnessax oh my mistake, I thought you meant your name was Xnessax :D hehehe [3:50 PM]

macclark52: Ha!  Color by positional entropy!   [3:50 PM]

Xnessax: haha [3:50 PM]

Nando: that's an option in the other graphs, yes [3:51 PM]

Xnessax: so its vanessa and Xnessax was inspirated by my cousin calling it as XnessaxHtinhaX  [3:50 PM]

Elves: Nando thanks for including the example link with preloaded sequence :D :D :D [3:51 PM]

tomoedachii: hey [3:51 PM]

Nando: but I'm talking of the bottom-most graph, mac [3:51 PM]

Elves: hi Tomoe!! :)  [3:51 PM]

Nando: hi Tomoe [3:51 PM]

tomoedachii: heyyy [3:51 PM]

macclark52: Ok, it IS positional entropy!  How cool.  [3:52 PM]

Xnessax: Hii tome...! [3:52 PM]

Elves: ohhhkay wow Nando this is a huge help [3:52 PM]

Nando: keep in mind guys, all of these predictions are only as good as the model (and its parameters) can be... [3:53 PM]

Elves: thanks for the reminder - this is algorithmic or in silico output, not direct data from lab experiments? [3:54 PM]

Nando: precisely [3:54 PM]

Xnessax: RNA and free enery? [3:55 PM]

Xnessax: energy* [3:55 PM]

Elves: and so i'm assuming the high entropy bulge / loop / junction here http://prntscr.com/92hvjh correponds to the peak of high uncertainty at that nucleotide position in the graph you mentioned? http://prntscr.com/92hw5k [3:56 PM]

Nando: yep, elves [3:57 PM]

Elves: so it is "positional entropy" in that each nucleotide position's potential entropy is predicted by the algorithm, and then an overall picture of the whole molecule is made by adding together each position [3:57 PM]

Xnessax: well i am lost in this dev [3:57 PM]

Xnessax: >< [3:57 PM]

Nando: not a simple matter of just adding actually, but close [3:58 PM]

Elves: Xnessax don't worry, I'll post the whole thing so you can see from the beginning [3:58 PM]

Elves: thanks for clarifying. i was wondering actually if adding was an appropriate term [3:58 PM]

Elves: i meant more combining, but I don't know :D [3:59 PM]

Nando: it's more like a combining, yes [3:59 PM]

Nando: as you can see in this example, ensemble diversity is pretty low [4:00 PM]

Elves: Xnessax the takeaway lesson I think is that when you make an RNA molecule in the labs, if you want it to be able to change easily into many shapes, then it helps if there are more options for each nucleotide to bind to [4:00 PM]

Nando: it's a single state target, and the EternaBot gives it a 95+ score, so it would be considered a rather good design [4:00 PM]

macclark52: So does that correspond to the fuzzy parts of the dot plot? [4:01 PM]

Nando: @mac: sort of, yes [4:01 PM]

macclark52: Sorry, I meant high positional entropy [4:01 PM]

Elves: mac Jen could explain best, but I think the fuzzy parts are points of partial probability for binding, and therefore if there is a lot of noise then the dot plot probably shows higher ensemble diversity [4:02 PM]

Nando: in both cases, ensemble diversity and positional entropy, you will "see" it as grey areas in the dot plot [4:02 PM]

Elves: i think the opacity or darkness of each dot corresponds to the level of probability of pairing. so light grey is low probability, dark or black is high, and nothing is no [4:03 PM]

macclark52: So positional entropy decreases as the P of the bond shifts away from .5 [4:03 PM]

macclark52: ? [4:04 PM]

Nando: yes [4:04 PM]

Nando: for every base, the less diverse option is to be either paired with nothing, or certainly paired with a specific base [4:05 PM]

Malcolm: Thank you Nando and Elves. This is one of the most interesting conversations I've seen on IRC. ;) :) [4:05 PM]

macclark52: Yes, you want a clear bias one way or the other.  [4:06 PM]

Nando: hi Malcolm, and glad you liked it :) [4:06 PM]

Elves: hehehe thanks Malcolm!!! that is awesome [4:06 PM]

Elves: glad to see you :) [4:06 PM]

macclark52: If I had seen this conversation when I first started hanging around EteRNA, I might have run away screaming.  :-) [4:07 PM]

Elves: it's been very educational for me as well. Thanks Nando for sharing so much helpful information! [4:06 PM]

Elves: ahahaha mac [4:07 PM]

Nando: @mac: correct, but as I said earlier, there may be cases where you don't want it to be "too perfect", specially with riboswitches... [4:07 PM]

Xnessax: Elves oh cool, got it. now it's an example for Lab. interesting. [4:06 PM]

macclark52: Yes, I appreciate the patience you all have in explaining.  [4:07 PM]

Xnessax: Does it mean a lesson for lab because if there's not tutorials for lab, right? [4:07 PM]

Xnessax: ah, got it, thanks Elves! [4:07 PM]

macclark52: There are going to be, I think.  [4:08 PM]

macclark52: So what might a good two state dot plot look like? [4:08 PM]

Elves: Xnessax exactly, we're taking every other Friday at Community chats to talk about things that will help us learn how to solve labs.  [4:08 PM]

macclark52: Mostly clean, but a little fuzz here and there?  [4:09 PM]

Nando: @mac: ah, if we only knew that :) [4:08 PM]

Elves: Ben Keep is working on tutorials to catch up with the progress labs has made, there are a lot of tutorials existing now but they are mostly for single state puzzles and basic switches [4:09 PM]

Elves: however, they can still help get up the beginning of the learning curve [4:09 PM]

macclark52: Do we know what the best energetic differential between states is? [4:09 PM]

Elves: and in the meantime, we're using the chats to learn more about terminology, strategies, etc. used in the Tuberculosis labs  [4:09 PM]

Nando: @mac: maybe that information is not enough... [4:09 PM]

macclark52: It's complicated.   [4:10 PM]

macclark52: Not as bad as protein folding maybe, but still complicated.  [4:10 PM]

Elves: Here's a link to chats with topics that may interest you http://eternawiki.org/wiki/index.php5/Dev_Chat [4:10 PM]

Nando: there's an area that we haven't really explored, because it's quite difficult: kinetics [4:10 PM]

Elves: here's a link to presentations from Das Lab, that help introduce these projects and concepts http://eternawiki.org/wiki/index.php5/Monthly_Eterna_Meeting [4:11 PM]

macclark52: Thanks, Elves.  [4:11 PM]

Nando: free energy barriers might be a more important factor than the differences in free energies between states [4:11 PM]

macclark52: So a big difference might signal a big energy barrier between states? [4:12 PM]

macclark52: Once you get down, you can't get back up.  [4:12 PM]

Nando: unfortunately, not in the general case [4:12 PM]

Nando: there's no way to infer the size of the barrier based only on the starting and ending points [4:12 PM]

Elves: And here is a very basic intro to tutorials and guides for learning skills to prepare, keeping in mind that the previous links are better for learning about current labs. http://eternawiki.org/wiki/index.php5/HELP [4:12 PM]

Nando: currently, you can only "sense" the barrier by running costly simulations called "trajectories" [4:13 PM]

Elves: ohhh hhahahaha [4:13 PM]

macclark52: This is sounding a little startrekky.  [4:14 PM]

macclark52: Thanks.  [4:14 PM]

Nando: http://rna.tbi.univie.ac.at/cgi-bin/barriers.cgi if you want to play with it [4:14 PM]

Elves: it sounds strangely similar to a problem in visualizing 3D objects with infinite resolution... :D [4:14 PM]

Nando: feed it a switch sequence, and see for yourself [4:14 PM]

Xnessax: Thanks Elves, i thought it was the objetive of every world wanting to understand about lab. [4:14 PM]

Xnessax: everyone* [4:14 PM]

macclark52: Bookmarked! [4:15 PM]

Elves: Xnessax, yes we are all ideally working towards understanding labs. the current labs are the result of many years of research, so there is a learning curve, and some basic steps that come first. jump in wherever you feel comfortable! [4:15 PM]

Elves: Nando thank you for the super cool resources [4:16 PM]

Nando: pleasure [4:16 PM]

macclark52: Ditto! [4:16 PM]

Nando: it's getting late here though, so if you'll excuse me, I shall retire :) [4:16 PM]

macclark52: Thanks for staying late.  [4:17 PM]

Elves: sleep well! thanks again :D  [4:17 PM]

macclark52: It was really helpful.  [4:17 PM]

Elves: time for me to go to work too, and post the chat. thanks everyone for your participation and questions, that made it awesome!! [4:17 PM]

Nando: bye everyone [4:17 PM]

Elves: bye Nando! [4:17 PM]

macclark52: Bye all.  [4:18 PM]

Elves: bye Mac, thanks :) [4:18 PM]

Xnessax: Everyone sleepin [4:22 PM]

Xnessax: 10pm here yet. [4:22 PM]

Xnessax: Thanks again Elves, it's going to be fun. I think Astromon is learning well for it. [4:23 PM]

Xnessax: and i haven't time to try many but i think i made mistake in other lab puzzles [4:23 PM]

Xnessax: without knowing it. So isn't it a problem? [4:23 PM]

Elves: happy to help! Glad you could come Xnessax. Don't worry about making mistakes, that is actually a good way to learn. the data that comes back from the lab always tells what does and doesn't work. so there's no way to know for sure without trying [4:24 PM]

Xnessax: No way to know for sure without trying.... [4:25 PM]

Elves: just try your best, learn a little something new as you can, and participate. the data from the lab will lead the way. :D [4:25 PM]

Elves: yep [4:26 PM]

Xnessax: And thanks again ,when I get time I would like to try it and study a little more about docs  [4:25 PM]

Xnessax: Then trying another again ,as like others submits as people did ,right? [4:26 PM]

tomoedachii: yep :) [4:26 PM]

Xnessax: :) [4:26 PM]

Elves: same here. there's more to know than any one person can figure out, so just study what interests you and try your best. [4:26 PM]

Xnessax: Indeed! [4:28 PM]