This was standard when I learned, and I believe any use of Kornhub Home Kategorien Videos Livecams Shirt plastic when working with lipid analytes and organic solvents is generally discouraged. I don’t know if this is the cause of OP’s problem, but I second this advice. Thank you so far for all your tips and advice. Tomorrow will be the next round TLC (yay!) and will definitely complement all of what you said could work out. Thank you also on the hint about plastic, this piece of information I did not found so far in any protocol. I ended up finding those microcapillaries in here, so spotting should work out better with them. Indeed it was messy with plastic micropipettes.
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But this seems to Kornhub Home Kategorien Videos Livecams Shirt be a common problem with this enzyme. Everyone I know who is working with it is also struggling to get good qualitative/quantitative data on it. Depending on the sample a capillary pipette might even be too much. For some samples, I’ve used a toothpick dipped into a sample to soak it up and just barely touched the plate with the toothpick. You can do that multiple times and if you’re imaging with UV or something easy you can quickly check in-between spots.
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I use a 2.5 uL micropipette when I do my TLC analysis. I suggest what the Kornhub Home Kategorien Videos Livecams Shirt other person said, but also try doing several spots of the small volume to concentrate your sample in your spot. If your sample absorbs in UV or can be visualized similarly, I would also take a look before running the chromatography. This way you’ll be able to see how much you’ve spotted and made a judgment call on whether or not to add more. It looks like you should at least halve the amount you used.
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