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    jsainton's Avatar
    jsainton Posts: 2, Reputation: 1
    New Member
     
    #1

    Sep 14, 2006, 09:41 AM
    real time PCR
    I am going to be doing a research study involving measuring the gene expression of chicken cytokines (IL-4 and IL-10) after being infected with a virus. I will be using real time PCR. However, I am confused as to the proper methods to use. Why is cloning necessary (creating plasmids)? Is this such an instance where I would have to cone? What real time PCR kit will work best for me? Roche, qiagen?

    Thank you very much.
    medgen's Avatar
    medgen Posts: 32, Reputation: 5
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    #2

    Nov 3, 2006, 12:58 PM
    1. No cloning is necessary -- you will be looking at mRNA. You only need to design appropriate primers

    2. TaqMan is by far the most specific and accurate method (Applied Biosystems). Sybr Green is your other option, but not nearly as specific.
    sovaira's Avatar
    sovaira Posts: 271, Reputation: 10
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    #3

    Nov 28, 2006, 03:48 AM
    Hello ,

    So you are going to begin research ,we ll in this field am a beginner ,well realtime PCR is abit complicated one ,well all PCRs r complicated in a way that you need to standardize the protocol,well Qiagen PCR kit may help,so wish you best of luck.

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