apricimo

I think this is my going out with fires of glory. This forum is filled with a bunch of people who don't have a clue about 90% of topics they post on. This includes YOU. Some of the "answers" you gave to my topics have been completely off base. Why chime in stuff you don't know. Why. People try to answer questions to topics they have no clue about. I'm done with it. Yes and thank you I know how committees are put together but again thank you for knowledge you put in that no one asked about. That seems to be your bread and butter. TO ANYONE WHO READS THIS BE ADVISED THIS FORUM IS WORTHLESS AND IS WORSE THAN GOOGLING AND READING A RANDOM WEBSITE. PEACE. "EXPERTS"

"General science advisor" on his thesis committee? What is general science? First of all what may that be. Secondly if you are on his committee in which case no one would refer to themselves as "general science advisor" why are you asking these things in science forums? Find related articles to confirm or disprove what he said. Were/are you working in the same as lab him and you don't believe what he has to say or just someone who knows this guy through random people? Thesis committee member asking questions about a thesis defense in science forums what a joker. Some people on here I swear. Start your own post about a question you have and don't self aggrandize yourself. General science advisor on a thesis committee.. My god Anyway to respond to the original post... It takes A LOT OF MONEY and A LOT OF WORK to do something that you read or heard about. You need fancy equipment and extremely bright scientists and those cost money. Bright scientists typically, though not always, would rather work on something legitimate since the amount of time they put in to acquiring their knowledge/degrees would be compromised/waste if they were doing something illegal. If there were/are people working on making super HIV strains I'm sure by now something would have leaked. That said there may be labs out there that may have found or are close to finding a way to alter the structure of HIV to a point where they can dictate function and we may not know about because such a discovery would be of interest to the forces of good and evil. Anything that is of high importance to humanity for reasons of good or bad is always a controversial topic and the internet a crappy source for it. So think rationally when you read stuff related to this and other important topics. Read peer reviewed journals and even then you have to be an honest skeptic. It takes A LOT OF MONEY and A LOT OF WORK to do something that you read or heard about. You need fancy equipment and extremely bright scientists and those cost money. Bright scientists typically, though not always, would rather work on something legitimate since the amount of time they put in to acquiring their knowledge/degrees would be compromised/waste if they were doing something illegal. If there were/are people working on making super HIV strains I'm sure by now something would have leaked. That said there may be labs out there that may have found or are close to finding a way to alter the structure of HIV to a point where they can dictate function and we may not know about because such a discovery would be of interest to the forces of good and evil. Anything that is of high importance to humanity for reasons of good or bad is always a controversial topic and the internet a crappy source for it. So think rationally when you read stuff related to this and other important topics. Read peer reviewed journals and even then you have to be an honest skeptic.

Looking to coexpress two vectors pCDFDuet and pETDuet in E. coli. Origins of replication are CDF and F1 respectively. Anyone know if these are compatible? Yes they are compatible

First Part: "Create" no but if you have the DNA sequence you can express it in e. coli or some other organism Second Part: Yes if you have the DNA sequence you can pretty much express any mutant you want. Doesn't mean it will be viable. On the other hand you can create a supermutant that has properties superior to the wild type Third Part: It is done in the lab all the time. Manually? Well if you are the one doing the work then it is manual I guess. No such thing as automatic work that you can do.... Comment: People who work with viruses typically make coat proteins to study. The genetic material for propagation of the virus is taken out so that you don't infect yourself.

All these types of "theories" start with something that has already evolved. For example, let's say that no theories on how life started exist as of right now, that is, no creationism or evolution or religion to cloud your thoughts. I come into the picture and say that life begins with water because water is needed for everything and continue on with this logic and maybe get two or three more "fundamental" essentials of life. But water is not fundamental, I used water because I see it and I am used to it but who says that water always "was" and whose to say what came from what. Best you can do is put the pieces together as you live your life and if they fit and everything you learned in life makes sense then maybe you are on the right path, but often times people are born into thinking a certain way. I come from a religious family and it took some time before I stopped feeling guilty that I didn't believe in God and creationism. Quite simply it doesn't make sense does it? Those that appose my view will say something along the lines of that all of this didn't come from nothing and that something had to create it, but I ask then how was that original something created. Typical response is that that something was always there and always will be. This is a dangerous circular logic game they play and because it is "self consistent" it seems valid. It is not, it is simply a subset of all truth whatever that truth may be I don't know but if you I do will let you know. Consider group theory where you have a special tupe of a group that is closed and all the elements that are added and multiplied generates another element that is already present in the group (D4h group in chemistry for example). Such a group is completely valid and often times useful but there are other groups which are more general that encapsulate the former group and others that encapsulate the former former and so on and so forth. The point is that knowledge should never be a perfect element where the end is also the start but rather something that is constantly evolving. An infinitely expanding sphere perhaps. Anyway, dimensional analysis is your first objective and I saw someone reply with no you cannot because the units come out weird. Yes dimensional analysis is always your first objective.

You lost me... looking at the character table I don't see a secondary axis.... that's fine my question was more of applying definitions in instances where they "don't apply" but I got it figured out...

Figured it out... In the absence of C2 perpendicular axis it applies to a vertical plane of symmetry... the labels make sense now Where is there i symmetry in C2v?

Ok question about the Mulliken symbols... In a C2v character table z is assigned a mulliken symbol A1. A is defined as a nondegerate and symmetric to principal axis and 1 is defined to mean symmetric with respect to C2 perpendicular to principal axis. Taking Water as an example for C2v symmetry which has no perpendicular C2 axis how do you assign the mulliken symbol A1. Also, same thing for y and x which are assigned B2 and B1 respectively. Don't get how these symbols come about when they don't apply to the definition of what they stand for. Please help....

Has anyone got gabedit 2.3.6 and ORCA 2.8 to work together? If so please let me know how... Gabedit seems to create the input file just fine but when called on ORCA through gabedit it seems to stall...

I am doing some cloning and using Blue/White screening and I was wondering if anyone has ever done this and ran into a problem of having all white colonies. I am using pGEM-T easy system for cloning. My background control is white also, but I don't understand why this would be.

excellent

I heard about a "rule of thumb" that you need your vector to be twice the size of the insert. So for a 10 kb fragment I would need a vector that is 20 kb. I can't think of any vectors like this and have you or anyone heard of this rule of thumb?

Hello, I am cloning a large, 10 kb, DNA fragment and I was wondering if anyone has had any experience with cloning large chunks of DNA. Background: The 10 kb DNA fragment has four genes in an operon. I would like to basically take this operon and express the proteins in E. coli so that they can do their thing (i.e. make the product that they make). Anyway, I heard different things from different people as far as what subcloning vectors to use or not use. For example, pGEM-T some people say use some people say that my insert is too large. TOPO vector I understand will work for something like this. My questions: 1.) What subcloning vector should I use for a fragment this large? 2.) What expression vector should I use to express my protein in E. coli with a fragment this large. The proteins are presumable cytosolic and come from a gram positive bacteria if that makes any difference in terms of expression vectors. I should make it clear that I am not trying to purify the proteins in the end rather I am trying to isolate the product that they make. Any help would be awesome. Thanks in advance

define topology with Spartan or with the server website? The energy minimisation in Spartan is on par with any other QM program out there (Gaussian).

So for example if I take a pdb of some protein from RCSB and I want to design a drug for it. HIV protease inhibitor, for example, in my case I am using AutodockTools with VINA so I need to have a pdb of the protein and the molecule I am docking also in pdb format. Hope that is clear I ended up using VMware fusion with Windows XP and ran Spartan with ChemDraw to draw my molecule and generate pdb style coordinates but it can be done also on a website called http://davapc1.bioch.dundee.ac.uk/prodrg/

I'll look into thanks. Sometimes it just takes knowing what to look for (extended range polymerases got me SuperTaq). I will the coolest microbiologist in the lab using SuperTaq

Hello I am trying to amplify and clone a 10 kb DNA fragment and I am currently using a Pfx platinum kit. I can get fragments <2 kb amplified no problem but I can't get the 10 kb product. Anyone know of a good system to amplify large chunks of DNA? I have a high GC organism I am working with also so I have to use enhancers (FYI I guess).

Why not? Anyway since posting this I figured out how to do all of it on a mac. So now you can't say that people ONLY use Windows and Linux for scientific work. Unix is a popular computing platform and that's what a mac is a UNIX.

Looking for a way to create a molecule and then spit out molecular coordinates in pdb format help please I should mention I am using a mac open source software is welcomed

ok there is no lac promoter

But it doesn't have a lac operon... What does IPTG do?

Is pET-3b used for expression? If so how do you induce it? If you don't induce it how does it work? If it is not an expression plasmid what is it used for? thanks

Seasnake.... Make a video with some type of screen recorded and a voice over and explain what you have in your equations and what you think they mean and their implications. Post it on you tube and let us know when you have done so. When people try to decipher your work and it is not in a published format people get cranky. It's easy to do.. For a mac screenflow works great... I don't know about windows...

The difference in taking the derivatives... Thought that was clear