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请各位帮忙设计RTPCR的引物,多谢了

丁香园论坛

1774
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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