帮忙看看我设计的引物

我用primer premier设计的引物,但是好像有错配.
1ctctccaaga agactcagcc agacccactc cagctccgac cctaggagac cgacctcctc
61 cagacggcag cagccccagc ccagtggaca accccaggag ccaccacctg gagcgtccgg
121 acaccaacct ccgccccgag accgagtcca ggctccggcc gcgcccctcg tcgcctctgc
181 accccgctgt gcgtcctcct gccgcgcccc gaccatgctc gcctccgtcg cgggtcccgt
241 tagcctcgcc ttggtgctcc tcctctgcac ccggcctgcc accggccagg actgcagcgc
301 gcagtgtcag tgcgcagctg aagcggcgcc gcgctgcccc gccggcgtga gcctggtgct
361 ggacggctgc ggctgctgcc gcgtctgcgc caagcagctg ggagaactgt gcacggagcg
421 tgatccctgc gacccacaca agggtctctt ctgcgacttc ggctcccccg ccaaccgcaa
481 gattggcgtg tgcactgcca aagatggtgc accctgtgtc ttcggtgggt ccgtgtaccg
541 cagcggcgag tccttccaaa gcagttgcaa ataccagtgc acttgcctgg atggggccgt
601 gggctgtgtg cccctgtgca gcatggacgt gcgcctgccc agccctgact gccccttccc
661 gagaagggtc aagctgcccg ggaaatgctg tgaggagtgg gtgtgtgatg agcccaagga
721 ccgcacagtg gttggccctg ccctagctgc ctaccgactg gaagacacat ttggccctga
781 cccaactatg atgcgagcca actgcctggt ccagaccaca gagtggagcg cctgttctaa
841 gacctgtggg atgggcatct ccacccgggt taccaatgac aataccttct gcaggctgga
901 gaagcagagt cgtctctgca tggtcaggcc ctgtgaagct gacctagagg aaaacattaa
961 ggtaaataaa tactctgccc ccagacactc attttcacag aatgacagga aagagaacca
1021 gagctggctg tctcgccgcc cttcttatta taggcctgtt gtctccagaa atatctaacc
1081 atggagctgt ctggctagaa tgagagatgc tgtaacagca gctgccagtt ttccactaca
1141 aaattccccg gggtgttagt taatacgaga cactccaaac gaagccatgg ctatttttgg
1201 aaaactggcg aatgaaactc ccagtctctc tcccctcaga atataaacac aagtcagatg
1261 acatagggct agtctacaga gggttgagga aggccactcc cgttgtagta attgcggtgt
1321 ctcttctcgt cttcccttag aagggcaaaa agtgcatccg gacgcctaaa attgccaagc
1381 ctgtcaagtt tgagctttct ggctgcacca gtgtgaagac ctaccgggct aagttctgtg
1441 gggtgtgcac ggacggccgc tgctgcacac cgcacagaac caccacactg ccggtggagt
1501 tcaagtgccc cgatggcgag atcatgaaaa agaacatgat gttcatcaag acctgtgcct
1561 gccattacaa ctgtcccggg gacaatgaca tctttgagtc cttgtactac aggaagatgt
1621 atggagacat ggcgtaaagc cagggagtaa gggacacgaa ctcatttaga ctataacttg
1681 aactgagtta catctcattt tcttctgtaa aaaaacaaaa aggattacag tagcacatta
1741 atttaaatct gggttcctaa ctgctgtggg agaaaacacc ccaccgaagt gagaaccgtg
1801 tgtcattgtc atgcaaatag cctgtcaatc tcagacactg gtttcgagac agtttagact
1861 tgacagttgt tcactagcgc acagtgacag aacgcacact aaggtgagcc tcctggaaga
1921 gtggagatgc caggagaaag acaggtacta gctgaggtca ttttaaaagc agcgatatgc
1981 ctactttttg gagtgtgaca ggggagggac attatagctt gcttgcagac agacctgctc
2041 tagcaagagc tgggtgtgtg tcctccactc ggtgaggctg aagccagcta ttctttcagt
2101 aagaacagca gtttcagcgc tgacattctg attccagtga cactggtcgg gagtcagaac
2161 cttgtctatt agactggaca gcttgtggca agtgaatttg ccggtaacaa gccagatttt
2221 tatggatctt gtaaatattg tggataaata tatatatttg tacagttatc taagttaatt
2281 taaagacgtt tgtgcctatt gttcttgttt taagtgcttt tggaattttt aaactgatag
2341 cctcaaactc caaacaccat cgataggaca taaagcttgt ctgtgattca aaacaaagga
2401 gatactgcag tggaaactgt aacctgagtg actgtctgtc agaacatatg gtacgtagac
2461 ggtaaagcaa tggatcagaa gtcagatttc tagtaggaaa tgtaaaatca ctgttggcga
2521 acaaatggcc tttattaaga aatggcttgc tcagggtaac tggtcagatt tccacgagga
2581 agtgtttgct gcttctttga ctatgactgg tttgggaggc agtttatttg ttgagagtgt
2641 gaccaaaagt tacatgtttg cacctttcta gttgaaaata aagtatatat attttttata
2701 tgaaaaaaaa aaaaaaaaaa aaaaaa
length Tm GC
S:ACCGC ACAGA ACCAC CACA 19 59.8 57.9
A:GAGAA GACGC TGAAG CCGA 19 57.9 57.9