Koch Lab:Protocols/Dig-bio PCR/pRL574 4411 PCR

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Contents

Materials

Primers

Forward Primer

5'-dig-GTAAAACGACGGCCAGTGAATTC (The Koch Lab calls this "pRL574-F853-dig")

  • Should hybridize to the forward strand starting at basepair 853

Reverse Primer

5'-bio-GGAAACAGCTATGACCATGATTAC (The Koch Lab calls this "pRL574-R5263-bio")

  • Should hyrbridize to the reverse strand starting at basepair 5263

Control Primers

It is not too expensive to purchase non-labeled primers to use as controls in case the PCR reaction doesn't work. Also sometimes it is convenient to perform PCR with only one label.

Primer Vendor

Steve endorses Alpha DNA for oligonucleotides. There are many other vendors--it's a good idea to go with a recommendation for a good digoxigenin-labeled oligo vendor.

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