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请帮我看看我设计的引物怎样 blast的结果可以吗

丁香园论坛

1982
要进行RT-PCR的序列就是 Oryctolagus cuniculus glucocorticoid receptor
1 atggactcca aagaatcatt aagtcccccg ggtagagagg aggtccccag cagtgtgctc
61 aggccggcgg agcgcgggaa tgtgatggac ttgtacaaaa ccctaagggg aggagctcct
121 gtgagggtgc ctgcatcttc tccctcactg gctcctgctg cgcagccgga ctccaagcag
181 cagagactcg cggtggactt tccgaaaggc tcagccagca atgcccagca gccggatctg
241 tccagagcag tgtcactctc gatgggattg tatatgggcg agacagaaac caaagtgatg
301 ggcagtgacc tggctttccc gcagcagggc caaaccagcc tttcctctgg ggaaacggac
361 ttccggcttc tggaagaaag catcgcgagc ctcaacaggt cggctagtgg tgcagacaac
421 cccaggagca cagcgcctgc tgctgggtct gccgccccca cagaggggtt tccaaaaact
481 cactctgacc tggcctccga acggcagaat ccgaagggcc agactggcgg cagcgccggc
541 agtgccaagt tgcaccccac agaccaaagc acctttgaca ttttgcagga tttggagttt
601 tctggatccc caagtaaaga caggagcgag agcccttgga ggtcagacct gttgatggac
661 gagaactgtc tgctttctcc tttggcggga gaagatgatc cattcctttt ggaagggaat
721 tcgagcgagg actgtaagcc tcttatttta ccggacacga aacctaaaat taaggataat
781 ggagatctga tcttatcaaa ctccaacaat gtaccactgc cccaagtgaa aaccgagaaa
841 gaagatttca tcgaactctg cacccctggg gtaattaagc aagagaaact gggcccagtg
901 tactgtcagg caagcttttc tggggcaaat ataattggta ataaaatatc tgccatttct
961 gttcatggcg tgagtacctc cggaggacag atgtaccact atgacatgaa cgcacagcag
1021 caggagcaga agcctctttt taatgtcatt ccgccaattc ctgttggttc agaaaattgg
1081 aatcgctgcc aagggtctgg agatgacaac ttgacttccc tggggactat gaacttccct
1141 ggccggtcgg tgttttccaa cggctattca agtccaggaa tgagaccaga tgtgagctct
1201 cctccatcta actccacgac agcagcagga ccacctccca aactctgctt ggtgtgctct
1261 gatgaagcct caggatgtca ctacggggtc ttgacttgtg gaagctgcaa agtattcttc
1321 aaaagggcag tgaaaggaca gcacaattat ctttgtgctg gaagaaatga ctgtatcatt
1381 gataaaattc gaagaaaaaa ttgcccagca tgccgataca gaaaatgtct tcaagctgga
1441 atgaacttgg aagcacgaaa aacaaagaaa aaaataaaag gaattcagca gaccagcaca
1501 ggagtttcgc aagaaacttc tgaaaatcct tctaacagaa cagtagttcc tgcagcatta
1561 ccacaactca ccccaacact ggtgtcactg ctggaggtca ttgaacctga agtgttgtat
1621 gcagggtatg acagctcggt tccagactct acttggagga tcatgaccac actcaacatg
1681 ttaggtgggc ggcaagtgat tgcagcagtg aaatgggcaa aggcaatacc aggattcagg
1741 aacttgcacc tggatgacca aatgaccctc ctgcaatact catggatgtt tctcatggca
1801 tttgccctgg ggtggagatc atataaacaa tccagtggaa acatgctatg ttttgctcct
1861 gatctggtta ttaatgagca gagaatgact ctaccctaca tgtatgacca atgtaaacat
1921 atgctgtttg tttcctctga gttaaaacgg cttcaggtgt cttacgaaga atatctctgt
1981 atgaaaacct tactgcttct ctctacagtt cctaaagaag gtttgaagag ccaagagcta
2041 tttgatgaaa tccgaatgac gtacatcaaa gagctaggaa aagccattgt caagagggaa
2101 ggaaactcca gtcagaactg gcagcgcttt tatcaactga caaaactttt ggactctatg
2161 catgaagtgg ttgaaaatct ccttcactat tgcttccaaa catttttgga taagacgatg
2221 agtattgaat tcccagagat gctagctgaa atcatcacca atcagatacc aaaatattca
2281 aatggaaata tcaaaaaact tctgtttcat cagaaatga
//
我设计的引物
sense
GCAGCATTACCACAACTCACC
antisense
TTGCCTTTGCCCATTTCA

进行blast后得到如下结果
Score E
Sequences producing significant alignments: (bits) Value

gi|26000752|gb|AY161275.1| Oryctolagus cuniculus glucocorti... 42 4e-05
gi|998894|gb|S78179.1|S78179 Oryctolagus cuniculus type XII... 32 0.037
gi|31072033|gb|AC122153.4| Oryctolagus cuniculus clone LB1-... 28 0.58
gi|1496|emb|X52496.1|OCCAATP Rabbit mRNA for calcium ATPase 28 0.58
gi|1468|emb|X02814.1|OCATPDR Rabbit mRNA for Ca2+-ATPase 28 0.58
gi|510177|emb|Z34886.1|OCHEAMYO1 O.cuniculus mRNA for beta ... 28 0.58
gi|165536|gb|K02444.1|RABMYSBC Rabbit cardiac beta-myosin h... 28 0.58
gi|164738|gb|J04703.1|RABACATP Rabbit smooth muscle sarco(e... 28 0.58
gi|37574168|gb|AC123541.3| Oryctolagus cuniculus clone LB1-... 26 2.3
gi|31339879|gb|AC123540.3| Oryctolagus cuniculus clone LB1-... 26 2.3
gi|25137556|gb|AF474181.1| Oryctolagus cuniculus fibroblast... 26 2.3
gi|2677621|emb|Y08030.1|OCART2 O.cuniculus ART2 gene 26 2.3
gi|40218214|gb|AY495827.1| Oryctolagus cuniculus clone 179L... 26 2.3
gi|165006|gb|M91005.1|RABELAMB Rabbit endothelial leukocyte... 26 2.3
gi|32362343|gb|AC125503.3| Oryctolagus cuniculus clone LB1-... 24 9.0
gi|30959399|gb|AY217091.1| Oryctolagus cuniculus immunoglob... 24 9.0
gi|2407751|gb|AF019284.1| Oryctolagus cuniculus Ig heavy ch... 24 9.0
gi|29029251|gb|AC122152.3| Oryctolagus cuniculus clone LB1-... 24 9.0
gi|27373656|gb|AY171867.1| Oryctolagus cuniculus clone mp12... 24 9.0
gi|27311263|gb|AC121066.3| Oryctolagus cuniculus clone LB1-... 24 9.0
gi|14719261|gb|AF389372.1| Oryctolagus cuniculus microsatel... 24 9.0
gi|34809229|gb|AY386696.1| Oryctolagus cuniculus clone 27N5... 24 9.0
gi|34809228|gb|AY386695.1| Oryctolagus cuniculus clone 219D... 24 9.0
gi|5733977|gb|AF120276.1|AF120276 Oryctolagus cuniculus sar... 24 9.0
gi|165562|gb|M20014.1|RABNPKC Rabbit protein kinase C-n (nP... 24 9.0
gi|165097|gb|M32030.1|RABHFMO Rabbit hepatic flavin-contain... 24 9.0
gi|6579182|dbj|AB035707.1| Oryctolagus cuniculus ITI HC1 mR... 24 9.0
gi|38565530|gb|AY453841.1| Oryctolagus cuniculus atherin mR... 22 36
gi|34787424|gb|AC144399.2| Oryctolagus cuniculus clone LBNL... 22 36
gi|30348994|gb|AC124042.3| Oryctolagus cuniculus clone LB1-... 22 36
gi|30315002|gb|AF498015.1| Oryctolagus cuniculus clone A37 ... 22 36
gi|30315000|gb|AF498013.1| Oryctolagus cuniculus clone A02 ... 22 36
gi|971173|gb|U20763.1|OCFIBRA1 Oryctolagus cuniculus fibrin... 22 36
gi|21913259|gb|AF480925.1| Oryctolagus cuniculus clone A ra... 22 36
gi|29135572|gb|AC124040.3| Oryctolagus cuniculus clone LB1-... 22 36
gi|12003422|gb|AF212147.1| Oryctolagus cuniculus skeletal m... 22 36
gi|28173024|gb|AF515800.1| Oryctolagus cuniculus endogenous... 22 36
gi|18409575|gb|AY072063.1| Oryctolagus cuniculus inward rec... 22 36
gi|538244|dbj|D16544.1|RABOPN Oryctolagus cuniculus opn mRN... 22 36
gi|17243824|gb|AF407578.2|AF407578 Oryctolagus cuniculus ac... 22 36
gi|2304973|gb|U85094.1|OCU85094 Oryctolagus cuniculus induc... 22 36
gi|34809230|gb|AY386697.1| Oryctolagus cuniculus clone 225P... 22 36
gi|34809227|gb|AY386694.1| Oryctolagus cuniculus clone 38A2... 22 36
gi|5759113|gb|AF170493.1|AF170493 Oryctolagus cuniculus put... 22 36
gi|45758469|gb|AY552542.1|AY552537S6 Oryctolagus cuniculus ... 22 36
gi|45758464|gb|AY552537.1|AY552537S1 Oryctolagus cuniculus ... 22 36
gi|4039111|gb|AF077828.1|OCINMT2 Oryctolagus cuniculus indo... 22 36
gi|4039108|gb|AF077826.1|AF077826 Oryctolagus cuniculus ind... 22 36
gi|3930411|gb|AF058603.1|AF058603 Oryctolagus cuniculus clo... 22 36
gi|3789965|gb|AF095465.1|AF095465 Oryctolagus cuniculus fib... 22 36
gi|1150914|gb|U44055.1|OCKCASN2 Oryctolagus cuniculus kappa... 22 36
gi|463998|gb|U07549.1|OCNKCC2F Oryctolagus cuniculus kidney... 22 36
gi|463994|gb|U07547.1|OCNKCC2A Oryctolagus cuniculus kidney... 22 36
gi|687678|gb|U20055.1|OCU20055 Oryctolagus cuniculus comple... 22 36
gi|456375|gb|U03455.1|OCU03455 Oryctolagus cuniculus phosph... 22 36
gi|2736306|gb|AF031619.1|OCTF09 Oryctolagus cuniculus trans... 22 36
gi|12003426|gb|AF212149.1|AF212149 Oryctolagus cuniculus sk... 22 36
gi|12003424|gb|AF212148.1|AF212148 Oryctolagus cuniculus sk... 22 36
gi|1627|emb|X07790.1|OCMTI Rabbit MT-I gene for metallothio... 22 36
gi|2342651|gb|AF015252.1|AF015252 Oryctolagus cuniculus gro... 22 36
gi|8648992|emb|AJ291317.1|OCU291317 Oryctolagus cuniculus p... 22 36
gi|1279622|emb|X79928.1|OCSMMHC O.cuniculus gene for smooth... 22 36
gi|1606|emb|Z18243.1|OCKAPCAS Oryctolagus cuniculus encodin... 22 36
gi|395193|emb|X68650.1|OCRYARA O.cuniculus mRNA for ryanodi... 22 36
gi|463996|gb|U07548.1|OCNKCC2B Oryctolagus cuniculus kidney... 22 36
gi|1245375|gb|U50465.1|OCU50465 Oryctolagus cuniculus cardi... 22 36
gi|165715|gb|M14577.1|RABTCBCA1 Rabbit T-cell receptor germ... 22 36
gi|632491|gb|U15514.1|OCU15514 Oryctolagus cuniculus smooth... 22 36
gi|165082|gb|K03256.1|RABHBBB1B1 Rabbit beta1-globin gene (... 22 36
gi|165065|gb|J00664.1|RABHBB4 rabbit embryonic beta-4-globi... 22 36
gi|165024|gb|M96556.1|RABFLAPA Oryctolagus cuniculus 5-lipo... 22 36
gi|404774|gb|L22090.1|RABEIF4G Rabbit initiation factor 4-g... 22 36
gi|164988|gb|L08449.1|RABDIANMON Rabbit dimethylanaline mon... 22 36
gi|164908|gb|M14955.1|RABCT450G Rabbit cytochrome P-450Bc2 ... 22 36
gi|164831|gb|M59743.1|RABCA2RE Rabbit cardiac muscle Ca-2+ ... 22 36
gi|164735|gb|M93344.1|RABAA1AG Rabbit acid alpha-1 acid gly... 22 36
gi|164729|gb|M20015.1|RAB015 Rabbit, metallothionein-I, exo... 22 36
gi|217735|dbj|D11411.1|RABOSP Oryctolagus cuniculus mRNA fo... 22 36
gi|165800|gb|L01083.1|RABUGT2BC Oryctolagus cuniculus UDP-g... 22 36
gi|165798|gb|L01082.1|RABUGT2BB Oryctolagus cuniculus UDP-g... 22 36
gi|164790|gb|M18818.1|RABBGLOB Rabbit beta-like globin gene... 22 36
gi|33312416|gb|AF421948.1| Oryctolagus cuniculus strain AX/... 20 141
gi|37682327|gb|AY422766.1| Oryctolagus cuniculus haplotype ... 20 141
gi|37682321|gb|AY422763.1| Oryctolagus cuniculus haplotype ... 20 141
gi|37682311|gb|AY422758.1| Oryctolagus cuniculus haplotype ... 20 141
gi|34419659|gb|AC124041.3| Oryctolagus cuniculus clone LB1-... 20 141
gi|20384667|gb|AY029517.1| Oryctolagus cuniculus urokinase-... 20 141
gi|559966|gb|U13689.1|OCALREDS2 Oryctolagus cuniculus aldos... 20 141
gi|22074827|gb|AY122285.1| Oryctolagus cuniculus urokinase-... 20 141
gi|21913263|gb|AF480926.1| Oryctolagus cuniculus clone B ra... 20 141
gi|21262338|gb|AF434336.1| Oryctolagus cuniculus RIa-14b-H ... 20 141
gi|28415761|gb|AY227450.1| Oryctolagus cuniculus corneal hy... 20 141
gi|294475|gb|L12405.1|RABCUZNSOD Rabbit copper/zinc-superox... 20 141
gi|6692994|gb|M14068.2|RABIGKC02 Rabbit Ig germline kappa-1... 20 141
gi|847859|gb|U26341.1|OCU26341 Oryctolagus cuniculus Na and... 20 141
gi|3293006|emb|AJ001588.1|OCAJ1588 Oryctolagus cuniculus co... 20 141
gi|2444021|gb|U72742.1|OCU72742 Oryctolagus cuniculus UDP-g... 20 141
gi|2305229|gb|AF011902.1|AF011902 Oryctolagus cuniculus UDP... 20 141
gi|1666775|gb|U62242.1|OCU62242 Oryctolagus cuniculus immun... 20 141
gi|1666773|gb|U62241.1|OCU62241 Oryctolagus cuniculus immun... 20 141

那个scores和E value 到底是什么意思
我这个引物可以用吗,

>gi|26000752|gb|AY161275.1| Oryctolagus cuniculus glucocorticoid receptor alpha mRNA, complete
cds
Length = 2319

Score = 42.1 bits (21), Expect = 4e-05
Identities = 21/21 (100%)
Strand = Plus / Plus


Query: 1 gcagcattaccacaactcacc 21
|||||||||||||||||||||
Sbjct: 1552 gcagcattaccacaactcacc 1572

Score = 36.2 bits (18), Expect = 0.002
Identities = 18/18 (100%)
Strand = Plus / Minus


Query: 22 ttgcctttgcccatttca 39
||||||||||||||||||
Sbjct: 1726 ttgcctttgcccatttca 1709

>gi|998894|gb|S78179.1|S78179 Oryctolagus cuniculus type XII collagen mRNA, partial cds
Length = 1920

Score = 32.2 bits (16), Expect = 0.037
Identities = 16/16 (100%)
Strand = Plus / Plus


Query: 19 accttgcctttgccca 34
||||||||||||||||
Sbjct: 917 accttgcctttgccca 932

这个identities=16/16 又是什么意思,怎么都是100%啊
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