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        请各位帮忙设计RTPCR的引物,多谢了

        丁香园论坛

        1946
        各位大侠,我想用RTPCR方法来做水稻HSP70基因的表达,但是水稻的HSP70很多不知道选哪个好,就选了这个,另外我用这个基因的CDNA做了BLAST后发现其500BP和1900bp之间和其它的HSP70基因是相同的,我想设计个500BP和1900bp之间的引物就可以把所有的HSP70基因的表达鉴定出来了,是这样吗但是不知道如何来选择设计引物,哪位帮忙来设计一下

        AK065431. Oryza sativa (jap...[gi:32975449] Links

        LOCUS AK065431 2380 bp mRNA linear PLN 24-JUL-2003
        DEFINITION Oryza sativa (japonica cultivar-group) cDNA clone:J013021C17, full
        insert sequence.
        ACCESSION AK065431
        VERSION AK065431.1 GI:32975449
        KEYWORDS FLI_CDNA; CAP trapper.
        SOURCE Oryza sativa (japonica cultivar-group)
        ORGANISM Oryza sativa (japonica cultivar-group)
        Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta;
        Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae;
        Ehrhartoideae; Oryzeae; Oryza.
        REFERENCE 1
        AUTHORS The Rice Full-Length cDNA Consortium, National Institute of
        Agrobiological Sciences Rice Full-Length cDNA Project Team:,
        Kikuchi,S., Satoh,K., Nagata,T., Kawagashira,N., Doi,K.,
        Kishimoto,N., Yazaki,J., Ishikawa,M., Yamada,H., Ooka,H., Hotta,I.,
        Kojima,K., Namiki,T., Ohneda,E., Yahagi,W., Suzuki,K., Li,C.,
        Ohtsuki,K., Shishiki,T., Foundation of Advancement of International
        Science Genome Sequencing & Analysis Group:, Otomo,Y., Murakami,K.,
        Iida,Y., Sugano,S., Fujimura,T., Suzuki,Y., Tsunoda,Y.,
        Kurosaki,T., Kodama,T., Masuda,H., Kobayashi,M., Xie,Q., Lu,M.,
        Narikawa,R., Sugiyama,A., Mizuno,K., Yokomizo,S., Niikura,J.,
        Ikeda,R., Ishibiki,J., Kawamata,M., Yoshimura,A., Miura,J.,
        Kusumegi,T., Oka,M., Ryu,R., Ueda,M., Matsubara,K., RIKEN:,
        Kawai,J., Carninci,P., Adachi,J., Aizawa,K., Arakawa,T., Fukuda,S.,
        Hara,A., Hashidume,W., Hayatsu,N., Imotani,K., Ishii,Y., Itoh,M.,
        Kagawa,I., Kondo,S., Konno,H., Miyazaki,A., Osato,N., Ota,Y.,
        Saito,R., Sasaki,D., Sato,K., Shibata,K., Shinagawa,A., Shiraki,T.,
        Yoshino,M. and Hayashizaki,Y.
        TITLE Collection, mapping, and annotation of over 28,000 cDNA clones from
        japonica rice
        JOURNAL Science 301 (5631), 376-379 (2003)
        MEDLINE 22752273
        PUBMED 12869764
        REFERENCE 2 (bases 1 to 2380)
        AUTHORS Adachi,J., Aizawa,K., Akimura,T., Arakawa,T., Carninci,P., Doi,K.,
        Fujimura,T., Fukuda,S., Hanagaki,T., Hara,A., Hashizume,W.,
        Hayashida,K., Hayashizaki,Y., Hayatsu,N., Hiramoto,K., Hiraoka,T.,
        Hori,F., Hotta,I., Iida,J., Iida,Y., Ikeda,R., Imamura,K.,
        Imotani,K., Ishibiki,J., Ishii,Y., Ishikawa,M., Itoh,M., Kagawa,I.,
        Kanagawa,S., Katoh,H., Kawagashira,N., Kawai,J., Kawamata,M.,
        Kikuchi,S., Kishikawa-Hirozane,T., Kishimoto,N., Kobayashi,M.,
        Kodama,T., Kojima,K., Kojima,Y., Kondo,S., Konno,H., Kouda,M.,
        Koya,S., Kurihara,C., Kurosaki,T., Kusumegi,T., Li,C., Lu,M.,
        Masuda,H., Matsubara,K., Matsuyama,T., Miura,J., Miyazaki,A.,
        Mizuno,K., Murakami,K., Murata,M., Nagata,T., Nakamura,M.,
        Namiki,T., Narikawa,R., Niikura,J., Nishi,K., Nomura,K.,
        Numasaki,R., Ohneda,E., Ohno,M., Ohtsuki,K., Oka,M., Ooka,H.,
        Osato,N., Ota,Y., Otomo,Y., Ryu,R., Saitoh,H., Sakai,C., Sakai,K.,
        Sakazume,N., Sano,H., Sasaki,D., Sato,K., Satoh,K., Shibata,K.,
        Shinagawa,A., Shiraki,T., Shishiki,T., Sogabe,Y., Sugano,S.,
        Sugiyama,A., Suzuki,K., Suzuki,Y., Tagami,M., Tagami-Takeda,Y.,
        Tagawa,A., Takahashi,F., Takaku-Akahira,S., Tanaka,T., Tomaru,A.,
        Toya,T., Tsunoda,Y., Ueda,M., Waki,K., Xie,Q., Yahagi,W.,
        Yamada,H., Yamamoto,M., Yasunishi,A., Yazaki,J., Yokomizo,S. and
        Yoshimura,A.
        TITLE Direct Submission
        JOURNAL Submitted (05-DEC-2001) Shoshi Kikuchi, National Institute of
        Agrobiological Sciences, Department of Molecular Genetics, Head of
        Laboratory of Gene Expression; 2-1-2 Kannondai, Tsukuba, Ibaraki
        305-8602, Japan (E-mail:skikuchi@nias.affrc.go.jp,
        Tel:81-29-838-7007, Fax:81-29-838-7007)
        COMMENT This clone is one of the 28K full-length cDNA clones from japonica
        rice.
        URL : http://cdna01.dna.affrc.go.jp/cDNA/
        NIAS Rice Full-Length cDNA Project Team: Kikuchi,S., Satoh,K.,
        Nagata,T., Kawagashira,N., Doi,K., Kishimoto,N., Yazaki,J.,
        Ishikawa,M., Yamada,H., Ooka,H., Hotta,I., Kojima,K., Namiki,T.,
        Ohneda,E., Yahagi,W., Suzuki,K., Li,C., Ohtsuki,K., Shishiki,T. and
        Yamamoto,M.
        FAIS Genome Sequencing & Analysis Group: Otomo,Y., Iida,Y.,
        Fujimura,T., Ikeda,R., Ishibiki,J., Kawamata,M., Kobayashi,M.,
        Kodama,T., Kurosaki,T., Kusumegi,T., Lu,M., Masuda,H., Miura,J.,
        Mizuno,K., Narikawa,R., Niikura,J., Oka,M., Ryu,R., Sugano,S.,
        Sugiyama,A., Suzuki,Y., Tsunoda,Y., Ueda,M., Xie,Q., Yokomizo,S.,
        Yoshimura,A., Matsubara,K. and Murakami,K.
        Genome Exploration Research Group in Riken Genomic Sciences Center
        and Genome Science Laboratory in Riken: Adachi,J., Aizawa,K.,
        Akimura,T., Arakawa,T., Carninci,P., Fukuda,S., Hanagaki,T.,
        Hara,A., Hashizume,W., Hayashida,K., Hayatsu,N., Hiramoto,K.,
        Hiraoka,T., Hori,F., Iida,J., Imamura,K., Imotani,K., Ishii,Y.,
        Itoh,M., Kagawa,I., Kanagawa,S., Katoh,H., Kawai,J.,
        Kishikawa-Hirozane,T., Kojima,Y., Kondo,S., Konno,H., Kouda,M.,
        Koya,S., Kurihara,C., Matsuyama,T., Miyazaki,A., Murata,M.,
        Nakamura,M., Nishi,K., Nomura,K., Numasaki,R., Ohno,M., Osato,N.,
        Ota,Y., Saitoh,H., Sakai,C., Sakai,K., Sakazume,N., Sano,H.,
        Sasaki,D., Sato,K., Shibata,K., Shinagawa,A., Shiraki,T.,
        Sogabe,Y., Tagami,M., Tagami-Takeda,Y., Tagawa,A., Takahashi,F.,
        Takaku-Akahira,S., Tanaka,T., Tomaru,A., Toya,T., Waki,K.,
        Yasunishi,A. and Hayashizaki,Y.
        FEATURES Location/Qualifiers
        source 1..2380
        /organism="Oryza sativa (japonica cultivar-group)"
        /mol_type="mRNA"
        /cultivar="Nipponbare"
        /db_xref="taxon:39947"
        /clone="J013021C17"
        ORIGIN
        1 gctctcccac tcttcctcct ccccaaaccc tagccgccgc cgccggcgta ctcgagagaa
        61 acatcagcat ccatggcggg caagggcgag ggtccggcca tcggcatcga ccttggcacg
        121 acctactcgt gcgtgggcgt ttggcagcac gaccgcgtgg agatcatcgc caacgaccag
        181 ggcaaccgca ccaccccctc ctacgtcggc ttcaccgact ccgagaggct catcggagat
        241 gctgccaaga accaggtcgc catgaacccc atcaacaccg tctttggtgc caagcgtctc
        301 attggcagga ggtttagcga tgcttctgtt cagagtgaca ttaagctctg gcccttcaag
        361 gtgattgctg gacctggtga caagcctatg attgttgtca agtacaaggg tgaggagaag
        421 cagtttgctg cagaagagat ctcctccatg gtcctcatca agatgcgtga gattgctgag
        481 gcctaccttg gcaccaccat caagaatgcc gttgtcactg ttcctgccta cttcaatgac
        541 tcccagaggc aggccaccaa ggatgctgga gtgattgctg gtctcaatgt catgcgtatc
        601 atcaacgagc cgactgctgc agctattgcc tatggtcttg ataagaaggc caccagcgtt
        661 ggtgagaaga atgtcctcat ctttgacctt ggaggtggta cctttgatgt ctccctcctt
        721 accattgagg agggtatctt tgaggtcaag gccacagctg gtgacaccca tcttggtggt
        781 gaagattttg acaatcgtat ggtcaaccac tttgtgcaag aattcaagag gaagaacaag
        841 aaggatatca ctggcaaccc cagggctctc aggaggttga ggacagcttg tgagagggca
        901 aagaggaccc tgtcctccac tgcccagacc accattgaga tcgattccct gtatgagggc
        961 atcgacttct actcaaccat cacccgtgcc aggtttgagg agctcaacat ggatctcttc
        1021 aggaagtgta tggagcctgt ggagaagtgc ctcagggatg ctaagatgga caagagctct
        1081 gttcatgatg ttgtccttgt tggtggctcc actaggatcc cgagggtgca gcagctcctg
        1141 caggatttct tcaacggcaa ggagctttgc aagaacatca acccagatga ggctgttgct
        1201 tatggtgctg ctgtccaggc tgccatcttg agtggtgagg gcaacgagaa ggtccaggac
        1261 ctcctcctgt tggatgttac ccctctctct ctcggtttgg agactgctgg tggtgtcatg
        1321 accgttttga tcccaaggaa caccaccatt cccaccaaga aggagcaggt cttctccacc
        1381 tactccgaca accagcctgg tgtgctcatc caggtttatg agggtgagag gaccaggaca
        1441 cgtgacaaca acctgctggg gaagtttgag ctctctggaa tccctcctgc tcccaggggt
        1501 gttccacaga tcactgtttg cttcgacatt gatgccaatg gtatcctgaa cgtgtctgct
        1561 gaggacaaga ccaccgggca gaagaacaag atcaccatta ccaacgacaa gggcaggctt
        1621 agcaaggagg agattgagaa gatggtccag gaggccgaga agtacaagtc agaggatgag
        1681 gagcacaaga agaaggtgga gtccaagaac gcgctggaga actacgccta caacatgcgc
        1741 aacaccatca aggatgagaa gatcgcctcg aagctcccgg cagcggacaa gaagaagatc
        1801 gaggatgcca tcgaccaggc catccagtgg ctggacggca accagctcgc tgaggctgat
        1861 gagttcgatg acaagatgaa ggagctggag ggcatctgca accccatcat cgccaagatg
        1921 taccagggcg ctggcgcgga catggccggc ggcatggacg aggacgatgc tcccccggct
        1981 ggcggcagcg gtgctggccc caagatcgag gaggtcgact aagcgcgaaa tttgctagtg
        2041 atccgcgctt caagttatct tattgcatta ttagtgtctc ttttatgttg tgttaaaaaa
        2101 ctttgctgct atgacttatg atgataccct tggtctgttt gggcagtcct tttgaattga
        2161 agaactgcgt tgggagctct atgagaacta tgaattgcgt gagaaattta attattgtgg
        2221 tgcttgcagc gcctattatt tggtctctgt cccacgttta ttatttgcat gtgcgcattg
        2281 tttgctagca attgctgttg tgcttctcag ccaacatgta aactgtttgg attgtttctg
        2341 atttttggat gcatttttta aagcggatag tgaaggacgg
        //
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