prabhubct

you mixed equal amounts of each but are they equal to the individual spots?

you'll notice that the shape of spot 1 and the size of spot 3 are altered, as well. the spots in the mixture are affecting the migration of each other. try the combinations of 2 at a time (1+2, 2+3, 1+3) and compare the effects.

Now I know what you mean.

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A Few Good Men in my lab.

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I guess.......still hanging.......

Thats just general information, but which bacteria you want to produce rabbit Ab. It will be take 3 month i guess in rabbit. Whole bacteria means just its capsule, isnt? You might look up the consistence of the capsule. It dosent matter if it Gram-negative or -positive, isnt? But i guess it will be unspecific Antibody. Do your Bacteria has specific marker molecules on his surface?

http://www.uniprot.org/keywords/875

http://ecvam.jrc.ec....hopReport35.pdf

Ah, that's a mycoplasma test. Mycoplasma don't usually kill your cells and the test that you linked has quite a low sensitivity.  It is quite likely (actually certain) that the cells are contaminated before they show up as positive.  Your best bet is to throw out your cells and probably any recent stocks and get some more in from a reliable source (e.g. ATCC).

You should also go through the incubators and give them a good clean, change your lab coat, throw out any opened stocks of medium, PBS, trypsin, and other reagents you are currently using.  Appraisal of your sterile technique would be a good thing to do as well, especially if you are new to cell culture.

If you have used BLAST search, you may find that repetitive sequence in your input is replaced by "n"s, this is called masking. So masked genome means genome with repetitive sequence excluded because repetitive sequences from deep seq data that match to more than one location are usually  ignored.

A couple of things to keep in mind:

• The height of the buffer in the chamber. Though you are putting in a specific voltage and amperage  more buffer will increase resistance, and therefore take away from the speed at which your bands run. Looking from the side of the box 0.5-1.0 cm above the cell is usually what I run. Not more. Plus, try to keep it consistent from gel-to-gel.
  
• The initial run speed. Most people ignore this, but it's a very useful trick. If I'm running a 120V gel, I always run it at 90V until the sample is all the way out of the well and into the lane (this is usually ~10 minutes). This way I can be more confident that my samples are running at the same velocity through the well when it counts. Basically, this is allowing your sample more time to snake its way into the pores of the gel, rather than cramming it in.
  
• Clean the electrodes in the box (that interface with buffer) with some diluted acetic acid or soap.
  
• This one is obvious, but make sure you're using 1X buffer.

If I think of others, I'll come back.

the glycine is a buffer that extends the useful range of the phosphate buffer with overlapping buffering ranges.

It should produce Abs, but the problem will be that some parts will be more antigenic than others and hence the antiserum might not be as useful as you would suspect.

Neuron,

if you immunize with a complex of proteins you will raise a polyclonal response. Given that some epitopes are immuno-dominant compared to others, most probably the result will be that you get antibodies to only a few of the proteins of your complex. I have done this several times with that results; there is also publications confirming this results. Hope this helps!

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Well are you really a newbie then with tech experience and a master's degree?
Anyway if the presentations are about topics that belong closely to your field of work, then you should work on theory and background and try to broaden your knowledge or try to catch up the stuff with a textbook.

If it's something different (belonging to different fields, nothing or not much to do with your work) then I think it's quite normal not to understand everything and even to have a quite relaxed view on this....detailed knowledge about this won't help you and you'll forget it anyway fast and biological sciences are extremely broad and you never can know all.
Best is perhaps to get a broader knowledge base and not to become an "expert idiot" with extremely restricted knowledge and view. And you might can try to learn more about basic principles and "laws" that often help to understand at least what they talk about, even if you don't get all tiny details.

And I guess in such classes most just pretend to understand everything and show the expert attitude...if you'd ask them to explain it, they'd be in difficulties

a good opportunity to use this good old website:
http://lmgtfy.com/?q...r dye chemistry