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I got problems

I got problems - PCR - Polymerase Chain Reaction Forum

I got problems - PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.


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  #1  
Old 03-09-2009, 05:20 PM
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Default I got problems



Hi all,
I tried to produce PCR product of mouse gene named SH3YL-1. I do not know what should I use as template. Or I have to buy it? Also I am so new for PCR area. I needs help.
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Old 03-10-2009, 02:30 AM
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Default Re: I got problems

Have you tried asking if anyone in your lab has the template? If yes (and if it is in the form of a plasmid), what you can do is to transform it into some competent cells to get more of the template, miniprep them and use them as template.
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  #3  
Old 03-10-2009, 03:27 AM
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Default Re: I got problems

are you trying to clone the cDNA? Usually I wouldn't do this (i previously thought it was a good idea working for several years in the lab), as PCR is error-prone and you may get a lot of amino acid changes if you are using it to express a protein.

You can always easily buy mouse cDNA clones in plasmids for cheap already cloned, and will ship to you.
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Old 03-10-2009, 08:50 PM
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Default Re: I got problems

Thank you for your replied. I asked everybody in my lab and they did not have it. So may be it was good idea to buy it as admin suggested. Or you have other ideas. Please Please
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