上下游引物GC%相差近20%,行吗?
丁香园论坛
3557
以下是primer3自动得到的引物,请高手指导,在线等。大写字母为外显子,小写的为内含子。
谢谢!!!!
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 501 20 61.99 55.00 6.00 2.00 AGCCTGGCCAACGTGTCTAT
RIGHT PRIMER 1026 22 59.42 36.36 2.00 0.00 tcgttgctgtttttctgttttc
SEQUENCE SIZE: 1250
INCLUDED REGION SIZE: 1250
PRODUCT SIZE: 526, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00
TARGETS (start, len~undefined: 521,430
481 GCTCTTCGCCAGCAACGTGAAGCCTGGCCAACGTGTCTATGTGCTGGGtGAGGGCGGGCA
>>>>>>>>>>>>>>>>>>>>~undefined******************~Kbr_~H~M~2~1~Kbr_~H~M~2~1541_GCAGCTGCTGCCGGCGTCTGTCCACAGCGTCTCATTGCGGGAGGAGGCGTCCGGAGCCTA~Kbr_~H~M~2~undefined**********************************************************~Kbr_~H~M~2~1~Kbr_~H~M~2~1601_CGCCCCACTCACCGCCCAGGGCACCATCCTCATCAACCGGGTGTTGGCCTCCTGCTACGC~Kbr_~H~M~2~undefined**********************************************************~Kbr_~H~M~2~1~Kbr_~H~M~2~1661_CGTCATCGAGGAGCACAGTTGGGCCCATTGGGCCTTCGCACCATTCCGCTTGGCTCAGGG~Kbr_~H~M~2~undefined**********************************************************~Kbr_~H~M~2~1~Kbr_~H~M~2~1721_GCTGCTGGCCGCCCTCTGCCCAGATGGGGCCATCCCTACTGCCGCCACCACCACCACTGG~Kbr_~H~M~2~undefined**********************************************************~Kbr_~H~M~2~1~Kbr_~H~M~2~1781_CATCCATTGGTACTCACGGCTCCTCTACCGCATCGGCAGCTGGGTGCTGGATGGTGACGC~Kbr_~H~M~2~undefined**********************************************************~Kbr_~H~M~2~1~Kbr_~H~M~2~1841_GCTGCATCCGCTGGGCATGGTGGCACCGGCCAGCTGAGAGTGGCTGGAGAGGGACAGAGA~Kbr_~H~M~2~undefined**********************************************************~Kbr_~H~M~2~1~Kbr_~H~M~2~1901_CAGAGTGTGGGAGAGAGAgagaaaaaaaaaaggggggggaggggggggggagaaagagaa~Kbr_~H~M~2~undefined************************************************~Kbr_~H~M~2~1~Kbr_~H~M~2~1961_aaaaaagaaaaaaaaaagaaaaaaagaaataacccaaatggaaagaaaacagaaaaacag~Kbr_~H~M~2~1~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~Kbr_~H~M~2~1~Kbr_~H~M~2~11021_caacgaaaaaaaaaaggaaaaataatacatatttttaaaagagagcacggattaagactt~Kbr_~H~M~2~1~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J
谢谢!!!!
OLIGO start len tm gc% any 3' seq
LEFT PRIMER 501 20 61.99 55.00 6.00 2.00 AGCCTGGCCAACGTGTCTAT
RIGHT PRIMER 1026 22 59.42 36.36 2.00 0.00 tcgttgctgtttttctgttttc
SEQUENCE SIZE: 1250
INCLUDED REGION SIZE: 1250
PRODUCT SIZE: 526, PAIR ANY COMPL: 5.00, PAIR 3' COMPL: 0.00
TARGETS (start, len~undefined: 521,430
481 GCTCTTCGCCAGCAACGTGAAGCCTGGCCAACGTGTCTATGTGCTGGGtGAGGGCGGGCA
>>>>>>>>>>>>>>>>>>>>~undefined******************~Kbr_~H~M~2~1~Kbr_~H~M~2~1541_GCAGCTGCTGCCGGCGTCTGTCCACAGCGTCTCATTGCGGGAGGAGGCGTCCGGAGCCTA~Kbr_~H~M~2~undefined**********************************************************~Kbr_~H~M~2~1~Kbr_~H~M~2~1601_CGCCCCACTCACCGCCCAGGGCACCATCCTCATCAACCGGGTGTTGGCCTCCTGCTACGC~Kbr_~H~M~2~undefined**********************************************************~Kbr_~H~M~2~1~Kbr_~H~M~2~1661_CGTCATCGAGGAGCACAGTTGGGCCCATTGGGCCTTCGCACCATTCCGCTTGGCTCAGGG~Kbr_~H~M~2~undefined**********************************************************~Kbr_~H~M~2~1~Kbr_~H~M~2~1721_GCTGCTGGCCGCCCTCTGCCCAGATGGGGCCATCCCTACTGCCGCCACCACCACCACTGG~Kbr_~H~M~2~undefined**********************************************************~Kbr_~H~M~2~1~Kbr_~H~M~2~1781_CATCCATTGGTACTCACGGCTCCTCTACCGCATCGGCAGCTGGGTGCTGGATGGTGACGC~Kbr_~H~M~2~undefined**********************************************************~Kbr_~H~M~2~1~Kbr_~H~M~2~1841_GCTGCATCCGCTGGGCATGGTGGCACCGGCCAGCTGAGAGTGGCTGGAGAGGGACAGAGA~Kbr_~H~M~2~undefined**********************************************************~Kbr_~H~M~2~1~Kbr_~H~M~2~1901_CAGAGTGTGGGAGAGAGAgagaaaaaaaaaaggggggggaggggggggggagaaagagaa~Kbr_~H~M~2~undefined************************************************~Kbr_~H~M~2~1~Kbr_~H~M~2~1961_aaaaaagaaaaaaaaaagaaaaaaagaaataacccaaatggaaagaaaacagaaaaacag~Kbr_~H~M~2~1~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~Kbr_~H~M~2~1~Kbr_~H~M~2~11021_caacgaaaaaaaaaaggaaaaataatacatatttttaaaagagagcacggattaagactt~Kbr_~H~M~2~1~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J~8lt~J