Dagl1

An overexpression screen? You have a WT yeast, in which you knockout an essential gene, now this has become a lethal phenotype, if during this time you now add start overexpression of a random gene, and your yeast survives, then you have rescued said phenotype. One way to do this is to make a overexpression library, get your plate/plates with knockouts and just add the library. Then pick the clones that survive and genotype them (or if you added barcodes to your plasmid library, identify by barcodes). If the essential gene knockout leads to a lethal phenotype only under certain conditions (cold sensitivity, some amino acid or other substance auxotrophicity) it is easier, because if your essential gene kills the yeast in all conditions, it is much harder to make sure your initial plate has 100% knockouts (since they won't grow). hope something like this helps

Could you not just design some primers for your specific genes, do PCR amplification and check on an agarose gel if the sizes are larger/smaller than expected? I am not entirely sure if that is the most feasible method, but at least I would assume you could see deletions, and duplications if they happened in the same region (of course if a duplication happened to be present in another chromosomal region, I would expect these to not be visible as the primers wouldn't lead to PCR amplification of 1 long transcript but just 2 equal sized ones, which I suppose you won't detect. Otherwise, can't you design some MLPA probes? I am not very familiar with qPCR MLPA validation, but if qPCR and sybr Green are used in this manner for duplication and deletion validation (even if it isn't the best way), then might just design the qPCR MLPA probe sets (I am not sure if that is feasible). I think you might find some information in: https://bmcmedgenet.biomedcentral.com/articles/10.1186/1471-2350-13-55 Maybe someone with more knowledge of this particular subject can come along and give you some more useful answers, but I think with that paper and other papers you should find some way ("I searched for deletion and duplication validation of novel genes"). -Dagl

I quoted only the relevant responses I want to discuss: 1. You say that each profile has its own specific pattern, and then you talk about gene expression and silencing, but that doesn't really answer my question: How did you determine what the boundaries are of 1 profile versus the next, if 2 people have the exact same gene expression except for 1 gene, are they the same profile? If so, at how much deviation do we find ourselves in another profile? How did you determine these boundaries? Have you considered alternatives? 2. So your profiles always match a trait, it never is wrong? What does 'to different degrees' mean when we are talking about matching. Either a profile associates with it or not right? Or have you scaled all the physical traits, so that one can get a score of 0 to x for 'long neckedness'? Please elaborate how they are matched, how things can be partially matched, and how is it possible that it matches always, you realise this is very unlikely with 26000 samples... 3. I am asking about how you chose to determine what physical phenotypes there were, you say it always matches, so how did you determine what a specific trait is. I don't care about why you chose these two to share, I am wondering about how you generated your list of total physical phenotypes, how you determined the boundaries and why not alternatives. 4. A list of people that I have to look up is not evidence, A I am a person, I can't quantify physical traits without significant amounts of bias, instead a computer should do this or a panel of people with very clear instruction. B Your list is going to be the definition of confirmation bias, I don't particularly doubt that these people from your dataset, for which you have produced epigenetic pattern definitions and physical trait definitions and which you then matched, will show the association that you are trying to tell us about. However, that doesn't mean anything; if I show you a bunch of pingpong balls that are all red, and I then tell you that I found red to be associated with pingpong balls, because, look all of the red pingpong balls that I am showing are red. Then this doesn't hold up. This is why we need frequency tables of the epigenetic patterns and of the physical traits, I would also love to actually see the statistics and the tests you have done to confirm your finding (again, you don't have to go showing everything, just of this one experiment). At the moment, I find it hard to believe that you have discovered anything, let alone came across an Nobel prize-winning discovery. This has little to do with the evidence, but a lot with your presentation and communication (skills). You might have discovered something really cool, but so far you have not provided evidence yet, and the lack in scientific rigour and the ease at which you apparently accept something to be evidence, makes it very hard for me to believe that you did actually find something. I think that regardless of what you have found, this is something you can improve on a lot! I am looking forward to hearing and seeing a bit more of the more methodical, quantitative and statistical side, since that is eventually how we can falsify information. Goodluck!

How are epigenetic profiles defined, how are they measured or determined, where are there boundaries? What does this list tell us? Is the frequency at which epigenetic profiles and these features match higher than what would be expected from regular distributions? How often do these profiles and features match and how often do they not? Are there differences for particular features or profiles? What is the total sample size of the evidence? How are these features defined, why these features and not other ones? At the moment, I don't really see any evidence yet, nor explanation of your methods. It would be very useful if you could describe in a few paragraphs what you have done (in this or another particular experiment, you don't have to explain ALL your evidence, just start by explaining a single one very well), and also describe the results. This txt file is literally a bunch of names, there seems to be no evidence yet, nor does it give me any real idea of what the list describes or what we should interpret from it. I hope you can provide more explanations and answer our questions! Kind regards, Dagl

This is a discussion forum, so people here would like to discuss things. You have stated you made some discoveries (and briefly and pretty vaguely described those discoveries), but what is there to discuss. I am interested in epigenetics, I would love to see and discuss your research, but then you need to post some of it. If you have so much evidence, what about sharing some of it HERE. Otherwise what is the point of this thread, without evidence there is nothing for us to discuss, so it seems like you are just (for a lack of a better term) gloating about discoveries made. Basically give us anything concrete, give us some evidence even if it is just a small part of all the discoveries you made. Or share the first 20(?) pages of your 'book' (not sure what to call it or how long it is). Things like 'causes of several epigenetic profiles' is pretty vague, it doesn't explain much and seems to be refuted by 'no', as that equals the amount of evidence provided. What tests did you to verify your discoveries, what alternatives exist to your explanation, did you test them. What about your general measurement methods, for what cells or organisms does this count? What is the difference between the epigenetic profiles? I assume you will have done some analyses of histone modifications, DNA methylation, residual RNA concentrations, and/or 3D genome structure. Just post any of those results, it could just be a bunch of pictures of Chip-seq analysis or whatever you have as evidence. Please note that I am not bashing you or your ideas, instead I hope to make you see why the current way of sharing your ideas may not be suitable for this forum. If you just want to let people know you made some discoveries, I think a profile message would be good. If you want to share your work, please do so! Me and others (probably) are interested in your discoveries, but with that must come the opportunity to review the evidence and doubt it. Right now it is similar to me saying that I have found a unifying theory of physics, or that I found novel protein functions, but that is all I tell anyone. Hope to hear more about your work soon! -Dagl

The specific type of lipid may signal that a cell is undergoing apoptosis. Under normal conditions specific lipids are moved to either the outside or the inside of the cell, during apoptosis (and maybe other cell death inductions) a scrambling protein is activated. You may be interesting in those enzymes: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4613456/ for the more general mechanism https://pubmed.ncbi.nlm.nih.gov/28844836/ additional links that I just browsed through: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4307283/ https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4914033/ Hope that helps!

A few questions and maybe some suggestions (that you need to verify and check, or use as step off points). I am not a statistician, but I do have experience with statistics. Hopefully I don't say anything just blatantly wrong;/ So you have k keywords, and their averages on a single day. Do you put k keywords into a single variable KEYWORD or do you want to measure whether there is a difference for every k keyword? If you want to do k comparisons, please apply some type of multiple comparison correction as your p value (assuming 0.05 is used as cut-off) only means the type of for a type 1 error is 5%, so if you compare 100 times, we need to make sure that that chance doesn't actually go up, and we do this by reducing the acceptable p value. You will have to provide some more information about your data: Is the variance constant? Is the average of your data a function of time? (so can you make an y= aT +b graph, if so the ARIMA I talk about later can't be applied I think (according to a video about it)) How long are the measurement periods, is seasonality included in your analysis? Now depending on these questions and some other assumptions (that I don't know but you will need to test), and what exactly you want to know (you seem to be interested in a change in frequency, so one could of course take the average frequency of a keyword in a similar period before and after the pandemic, and then just test the two group (each group will have n datapoints (amount of days of each period) and so you can calculate SEM, SD, 95%CI and do regular T-tests (as Prometheus has said, t-tests have assumptions such as the distribution of your data, you will have to test each of these assumptions with a separate test. If you do not meet all the assumptions you can use a non-parametric alternative of the t-test (one for which your data does have all the required things)). With this approach, you measure frequency over almost a year, so seasonality is something to think of. Taking datapoints from the same days in the previous year or years could of course be a way around this seasonality problem. If you do want to analyse your data as a time series, there are several things you need to consider: What exactly do you want to analyze. Are the time series 'different' is really dependent on what you are particularly interested in. Are you interested in the total frequency over time (AuC), are you interested in a change in frequencies per week (a shift from monday to friday), are you interested in something else? From a quick google search, ARIMA seems to be one approach, but I am not entirely sure how to apply that to two datasets. I suppose you would do ARIMA for both and see if they predict similar things, if not then they are different. But if this is the case, then you would still need to find a way to define when the two predictions are 'statistically different'. Maybe someone more into this part of analysis could help out here. https://stats.stackexchange.com/questions/35129/how-to-compare-two-time-series https://stats.stackexchange.com/questions/19103/how-to-statistically-compare-two-time-series Another thing I found is a fixed-effects ANOVA, it seems that the dataset is similar to yours if you can overlay the data from the same days before and after the pandemic (note that ANOVA generally are for 3 or more groups, which is the case on this website, so there most likely is a t-test-like variant of this that you want to use) https://stats.stackexchange.com/questions/12902/comparison-of-time-series-sets Although not that important at the moment, I do wonder if overlaying march 1 2019 and march 1 2020 is actually the best option (as days of weeks change with such an approach) and it may be better to shift it to match days of the week instead of date). I am not sure if that is actually better, but I think it may be an interesting thing to note in your discussion. I hope this at least helps you a bit, but it is important to check yourself what you are and are not allowed to do! A lot of tests that you can perform on SPSS have nice documentation somewhere on the internet that includes the assumptions which need to be met, I like this website a lot: https://statistics.laerd.com/spss-tutorials/linear-regression-using-spss-statistics.php) Kind regards, Dagl

@CharonY Thanks for the explanation! That adds quite a few things to the way I see it. Now back to the topic at hand (as I am also partly to blame for derailing it), do you all think that if this eventually dies down, and things do not change (in the system), these types of protests and riots will become more frequent, or will this be (at this scale) more a one-off thing?

Maybe I misunderstood, but you are saying tribalism and racism are not quite the same and I agree, but biologically speaking, we do have different responses (increased amygdala, reduced/slower prefrontal cortex inhibition when seeing out-group people) to in-group and out-group people. This is what I would consider tribalism, but one of the easiest ways for our brains to identify in- and out-group people is physical appearance (which is where racism starts, but I suppose maybe you mean that racism goes much further than such relatively fundamental responses)? Please correct me if I am wrong or misunderstood you, I also realise that studies such things as increased amygdala response when seeing people with a different skin-colour may fall in the non-reproducible category of psychology/neuroscience.

Together with your other statements, this answer seems as if you are not agreeing with the other points, or have not gotten any new knowledge or insight from peoples post, this could of course be completely untrue, but at the same time, based on the tendency of your previous answers and posts on other topics, it is not unreasonable (to me) to think that you still hold your initial believes, not understanding that systemic racism exists, denying its existence, or not seeing how the fact that it is currently a bigger issue for some people than for others (and should therefore first be solved for those people (referring back to the comment about every group of people (on average) possibly being equally as racist)). Some post ago you asked people in this forum to help you comprehend; they have attempted to, did it make a difference, do you now understand more than before? If not, why? Again I could be completely misreading it, but to me it feels like you evaded the other questions I asked.

Maybe it is an interesting thing to research yes, but does this, together with other people's explanation and Dimreepr's video explain to you the problem people (non-white/non-christian) are facing? Your previous posts seemed to ask why these things were problems, did these questions get answered and do you understand the problems and why people are currently battling racism coming from white people that is ingrained in the system (within the USA, and possibly other places in the western world)? Do you agree that white-people disproportionately hold power in society, and do you also agree with the fact (which is a strange question) that non-whites are more often victims of police misconduct, brutality, and assault. Do you think things should be systematically changed because of these things that have been shown to you?

You said it more clearly than I did!

Because some people have been on the receiving end of racism for much longer than others, potentially making their experiences different. But the problem is that currently white people hold much more societal power than those with other skin colours and those that are parts of religions that are not Christianity. This means that even if there would be equal racism among all people, the consequences disproportionately affect non-white/non-Christian (in the western world!) people. Hence people speak of systemic racism, yes it may be (please note that I am not saying this is the case) the case that if most power in the USA and other western countries would lie in people of colour, they would be equally racist. But they are not in power (generally) and thus the current issue is that they are disproportionally discriminated against. If we ever get to a position where it is the other away around, then we have a different issue to solve.

What things have you thought of, what things are things are there to consider in this question? What is osmosis and how does it work?

Would that be a bad thing? I do myself subscribe to the idea that those in power should be held to higher standards than those that are not, and that if you do something wrong or abuse your power, you should be punished more severely (the higher you are, the longer the fall. comes to mind).

I think Jack means that besides being punished for murder, the fact that this person was part of the system (society), and misused his power, he is breaking the social contract, which should be punished by life in prison with no chance of parole (according to Jack). If this police officer had not killed anyone, but still abused his power, I think Jack would also want this person to go to prison for life, without parole because it is breaking the social contract that individuals sign with (are forced to sign with) society.

How would you verify the localisation after you get some data? Anyway I hope this helps you on your way with your essay, I am signing off for now.

Sure! So we have a protein that is uncharacterized, meaning there probably are not going to be antibodies available. We can try producing proteins against it, but we could also try something else. What is cloning and what ability does it give us? How could we fuse these two things (fluorescence and cloning) together? The answer has a caveat though; there is of course a potential drawback to using fluorescence, in that it may not accurately describe localisation due to how we use fluorescence, but this is something that can be discussed/worked around later, and may not be in the scope of the question.

What are ways that we can find out the localisation of a protein? How do we employ this by using cloning?

I was thinking of free will in the sense of conditions (your actions now are the result of things that happened in the past, and the molecules in your body reacting to previous states, at no point do you have any 'say' in what the molecules do), not regarding the idea one could make some different choice based on reasoning, but I believe even with the definition that people can make meaningful choices, they are limited to whatever their personality/being/brainstate at that particular time allows, so that type of free will would be limited (but not non-existent). You confidently say what free will is, but I presume it is the definition (that I missed by not reading through the entire thread) you or people have agreed upon here, because I generally interpret free will in the sense as I described above. If we do follow my definition, I don't agree you would come into an endless loop when calculating everything, you would come to the conclusion that you calculating everything was part of what was/is going to happen, and it would all be part of some causal chain (of course, quantum mechanics and inherent randomness can make that chain deviate each time, but since humans (as far as I know) can't control the randomness of quantum mechanics, this deviation is still not in anyway influenced by some person). Regarding colour... well that is wavelengths of light being interpreted being picked upon our eyes right. Do electrons, protons, and neutrons not emit specific frequencies of light (even if they may not be within the visual spectrum)? (I honestly don't know, but always assumed they do). My point about the fact that calculating what YOU would do, would have to include myself into that equation, and to calculate what I would do, I would require my own environment, therefore me calculating whatever would be part of the causality, and therefore me telling you, and the specific reaction (you changing) would all be included in that calculation, thus you would have to change your calculation until the deviation would be so small that it is true. But I suppose it could also be an endless loop, depending if the act of knowing the calculations leads to divergent or convergent changes upon each subsequent calculation.

I don't entirely agree with this, because if I would predict what would happen based on the laws of physics, then the part where I am telling you, must be part of that prediction. Because the environment and all other molecules are part of that equation, and thus if I could predict what would happen, then I would have to include all the information that includes telling you, and thus it 'has' to happen. If I won't tell you, but then I would have predicted that I wouldn't tell you since I predicted it based on the laws of nature. It seems impossible to calculate ONLY your actions based on the laws of nature and present conditions, because for that I would have to calculate MY and the entire (local) environment, right? Your thoughts? (Hope I am clear, only have a few sec before I have to leave, otherwise I will reiterate later).

Do you understand the argument that free will is something you feel like you experience, but in no way can your actions prove that you were ever going to do something else. Yes you may say that you can reason and think about other options. but that reasoning/thinking could just as well be something you were gonna do regardless of your choice. Your brain may just make it seem like you have any control. If you understand those arguments, then you also understand how your example of posting on this forum, does nothing to refute that argument? Additionally, from a physical point of view, if you don't invoke a soul, then your brain is made of neurons, with proteins, all of which undergo chemical reactions that lead to thoughts/emotions (of course your body also has hormones, but those too are physical things). So if molecules don't have free will, how could you have free will. (My apologies if this is already talked about in this thread, I just skimmed the last pages).

Ah ye, fair point, thanks for the link!

You called it bias, but now there are links showing potential ties. Do you deny the validity of those claims or? Sure, iNow (I think) said this before evidence had accumulated, based on previous behaviour of the US president, is that bias? Or are you agreeing with everything, but just pointing out that people are biased (because bias doesn't mean people can't be right)? Could you elaborate? In my country (Netherlands), we have added it onto our recommended treatment options, I personally thought some trials were promising, and others weren't. There is no conclusive evidence so far, is there? (I think you are referring to the article Charon posted: No Evidence of Rapid Antiviral Clearance or Clinical Benefit with the Combination of Hydroxychloroquine and Azithromycin in Patients with Severe COVID-19 Infection)?

Music by 音阙诗听 like: But a lot of other Chinese/Taiwanese singers as well.