【心得】自己设计的三个基因的Qpcr的引物，供战友指正`讨论

因为是Qpcr引物，为了提高准确性，就缩短了产物长度，大概都在100-150bp左右。供高手讨论，共同提高。
目的基因一（CDS区）：
gacattgtctaaaagccaagatgacagactgagaggcctgagcccttgttctggcattctcccaggaagatgcagtaaaggggttgacccaatatactgcagagaatttcatccagttccctcctccatcctgatcccatgtgccaggaagatggagccacgcttgctgatgttggggtttctctcattaaccatagtacccagttgtcgggcagaactgtgtctgtatgacccacccgaggtccccaatgccacattcaaagccctctcctacaagaacggcaccatcctaaactgtgaatgcaagagaggtttccgaagactaaaggaattggtctatatgcgttgcttaggaaactcctggagcagcaactgccagtgcaccagcaactcccatgacaaatcgagaaagcaagttacagctcaacttgaacaccagaaagagcaacaaaccacaacagacatgcagaagccaacacagtctatgcaccaagagaaccttacaggtcactgcagggagccacctccttgggaacatgaagattccaagagaatctatcatttcgtggaaggacagagtgttcactacgagtgtattccgggatacaaggctctacagagaggtcctgctattagcatctgcaagatgaagtgtgggaaaacggggtggactcagccccagctcacatgtgtagatgaaagagaacaccaccgatttctggctagtgaggaatctcaaggaagcagaaattcttctcccgagagtgagacttcctgccccataaccaccacagacttcccacaacccacagaaacaactgcaatgacggagacatttgtgctcacaatggagtataaggtagcagtggccagctgcctcttcctgctcatcagcatcctcctcctgagcgggctcacctggcaacacagatggaggaagagcagaagaaccatctag
设计的引物：

upper:5`tgtctgtatgacccacc
lower:5`agtcttcggaaacctc

目的基因二（CDS区）：
agaccttggcagatgtgacttcagttcacaccacactctgccttgctcacagaggaggggctgcagccctggccctcatcagaacaatgacactcaggctgctgttcttggctctcaacttcttctcagttcaagtaacagaaaacaagattttggtaaagcagtcgcccctgcttgtggtagatagcaacgaggtcagcctcagctgcaggtattcctacaaccttctcgcaaaggaattccgggcatccctgtacaagggcgtgaacagcgacgtggaagtctgtgtcgggaatgggaattttacctatcagccccagtttcgctcgaatgccgagttcaactgcgacggggatttcgacaacgaaacagtgacgttccgtctctggaatctgcacgtcaatcacacagatatttacttctgcaaaattgagttcatgtaccctccgccttacctagacaacgagaggagcaatggaactattattcacataaaagagaaacatctttgtcatactcagtcatctcctaagctgttttgggcactggtcgtggttgctggagtcctgttttgttatggcttgctagtgacagtggctctttgtgttatctggacaaatagtagaaggaacagactccttcaaagtgactacatgaacatgactccccggaggcctgggctcactcgaaagccttaccagccctacgcccctgccagagactttgcagcgtaccgcccctgacagggacccctatccagaagcccgccggctggtacccgtctacctgctcatcatcactgctctggatagga

设计的引物：
upper:5`caggctgctgttcttg
lower:5`gctgacctcgttgctat

目的基因三（CDS区）：
ttctggagggcttctcttgcctgccccatgtcctgcaaatgcagcaccactaggatttggtgtaccgagccttctcctggcatcgtggcatttccgaggttggaacctaacagcattgacccagagaacatcaccgaaattctcattgcaaaccagaaaaggttagaaatcatcaatgaa1gatgatgtcgaagcttacgtggggctgaaaaaccttacaa4ttgtggattccggcttaaagtttgtggcttacaaggcgtttctgaagaacggcaacctgcggcacatcaatttcactcgaaacaagctgacgagtttgtccaggagacatttccgccaccttgacttgtctgacctgatcctgacgggtaatccgttcacgtgttcctgtgacatcatgtggctcaagactctccaggagacgaaatccagccccgacactcaggatttgtattgcctcaa2tgagagcagcaagaatacccctctggcgaacctgcagattcccaattgtggtctgccgtctgcacgtctggccgctcctaacctcacggtggaggaagggaagtctgtgaccatttcctgcagcgtcgggggtgacccgctccccaccttgtactgggacgttgggaatttggtttccaaacacatgaatgaaacaagccacacacagggctccttaaggataacaaacatttcatcggatgacagtgggaaacaaatctcttgtgtggcagaaaacctcgtcggagaa3gatcaagactctgtgaacctcactgtgcattttgcaccaaccatcacatttctcgaatctccaacctcagaccaccactggtgcatcccattcactgtgagaggcaaccccaagccagcacttcagtggttctacaacggagccatactgaatgaatccaagtacatctgtaccaaaatacacgtcaccaatcacacggagtaccacggctgcctccagctggataaccccactcatatgaataatggagactacaccctaatggccaagaatgaatatgggaaggacgag

设计的引物：
upper:5`agagcagcaagaataccc
lower:5`gacttcccttcctccacc

以上引物可能存在的共同问题：两条引物的3`都有引发合成的可能，导致假阳性率比较高。