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Usage:
after submit the task, the page will be automatically refreshed every 3 sec, please wait 2~5 seconds to see the updated result.
任务提交后,结果页面会每3秒自动刷新, 直到出结果;通常1-2秒出结果,如果是多重引物评估,一分钟之内能出结果。
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News:
Welcome to use our new website MFEprimer-3.1, which is faster and more effective.
Enter the primer sequences (multiplex PCR primers supported,
example
) *
Number of primer sequences: 1 (min) - 50 (max). 2 (max) primers for degenerate bases.
E-mail
For receiving the results. If you have more than two primers, we suggest you leave your E-mail. (Optional)
Background Database
UCSC - Homo sapien - hg19 - Genome
UCSC - Homo sapien - GRCh37(hg19) - Genome+RNA
UCSC - Homo sapien - GRCh37(hg19) - RNA
UCSC - Anopheles gambiae v. MOZ2 - anoGam1 - Genome
UCSC - Apteryx australis - aptMan1 - Genome
UCSC - Apis mellifera - apiMel1 - Genome
UCSC - Anolis carolinensis - anoCar1 - Genome
UCSC - Aplysia californica - aplCal1 - Genome
UCSC - Aquila chrysaetos canadensis - aquChr2 - Genome
UCSC - Anolis carolinensis - anoCar2 - Genome
UCSC - Apis mellifera - apiMel2 - Genome
UCSC - Balaenoptera acutorostrata scammoni - balAcu1 - Genome
UCSC - Bison bison - bisBis1 - Genome
UCSC - Bos taurus - bosTau2 - Genome
UCSC - Bos taurus - bosTau3 - Genome
UCSC - Bos taurus - bosTau4 - Genome
UCSC - Bos taurus - bosTau6 - Genome
UCSC - Bos taurus - bosTau7 - Genome
UCSC - Bos taurus - bosTau8 - Genome
UCSC - Branchiostoma floridae - braFlo1 - Genome
UCSC - Caenorhabditis japonica - caeJap1 - Genome
UCSC - Caenorhabditis brenneri (PB2801) - caePb1 - Genome
UCSC - Caenorhabditis brenneri (PB2801) - caePb2 - Genome
UCSC - Callorhinchus milii - calMil1 - Genome
UCSC - Caenorhabditis remanei - caeRem2 - Genome
UCSC - Caenorhabditis remanei - caeRem3 - Genome
UCSC - Ceratotherium simum - cerSim1 - Genome
UCSC - Chrysemys picta bellii - chrPic1 - Genome
UCSC - Caenorhabditis briggsae - cb3 - Genome
UCSC - Caenorhabditis elegans - ce10 - Genome
UCSC - Caenorhabditis elegans - ce11 - Genome
UCSC - Caenorhabditis elegans - ce2 - Genome
UCSC - Caenorhabditis elegans - ce4 - Genome
UCSC - Caenorhabditis elegans - ce6 - Genome
UCSC - Ciona intestinalis (sea squirt) - ci1 - Genome
UCSC - Ciona intestinalis (sea squirt) - ci2 - Genome
UCSC - Ciona intestinalis (sea squirt) - ci3 - Genome
UCSC - Chlorocebus sabaeus - chlSab2 - Genome
UCSC - Canis familiaris - canFam1 - Genome
UCSC - Canis familiaris - canFam3 - Genome
UCSC - Choloepus hoffmanni - choHof1 - Genome
UCSC - Cricetulus griseus - criGri1 - Genome
UCSC - Cricetulus griseus - criGriChoV1 - Genome
UCSC - Callithrix jacchus - calJac1 - Genome
UCSC - Callithrix jacchus - calJac3 - Genome
UCSC - Cavia porcellus - cavPor3 - Genome
UCSC - Dasypus novemcinctus - dasNov3 - Genome
UCSC - Drosophila melanogaster - dm1 - Genome
UCSC - Drosophila melanogaster - dm2 - Genome
UCSC - Drosophila melanogaster - dm3 - Genome
UCSC - Drosophila melanogaster - dm6 - Genome
UCSC - Drosophila pseudoobscura - dp2 - Genome
UCSC - Drosophila pseudoobscura - dp3 - Genome
UCSC - Drosophila ananassae - droAna1 - Genome
UCSC - Drosophila ananassae - droAna2 - Genome
UCSC - Drosophila erecta - droEre1 - Genome
UCSC - Drosophila grimshawi - droGri1 - Genome
UCSC - Drosophila mojavensis - droMoj1 - Genome
UCSC - Drosophila mojavensis - droMoj2 - Genome
UCSC - Drosophila persimilis - droPer1 - Genome
UCSC - Drosophila sechellia - droSec1 - Genome
UCSC - Drosophila simulans - droSim1 - Genome
UCSC - Drosophila virilis - droVir1 - Genome
UCSC - Drosophila virilis - droVir2 - Genome
UCSC - Drosophila yakuba - droYak1 - Genome
UCSC - Drosophila yakuba - droYak2 - Genome
UCSC - Dipodomys ordii - dipOrd1 - Genome
UCSC - Danio rerio - danRer10 - Genome
UCSC - Danio rerio - danRer11 - Genome
UCSC - Danio rerio - danRer3 - Genome
UCSC - Danio rerio - danRer4 - Genome
UCSC - Danio rerio - danRer5 - Genome
UCSC - Danio rerio - danRer6 - Genome
UCSC - Danio rerio - danRer7 - Genome
UCSC - Filoviridae ebolavirus - eboVir3 - Genome
UCSC - Echinops telfairi - echTel1 - Genome
UCSC - Echinops telfairi - echTel2 - Genome
UCSC - Equus caballus - equCab1 - Genome
UCSC - Equus caballus - equCab2 - Genome
UCSC - Erinaceus europaeus - eriEur1 - Genome
UCSC - Erinaceus europaeus - eriEur2 - Genome
UCSC - Felis catus - felCat3 - Genome
UCSC - Felis catus - felCat4 - Genome
UCSC - Felis catus - felCat5 - Genome
UCSC - Felis catus - felCat8 - Genome
UCSC - Takifugu rubripes v3.0 - fr1 - Genome
UCSC - Takifugu rubripes v4.0 - fr2 - Genome
UCSC - Takifugu rubripes v5.0 - fr3 - Genome
UCSC - Gadus morhua - gadMor1 - Genome
UCSC - Gallus gallus v1.0 - galGal2 - Genome
UCSC - Gallus gallus v2.1 - galGal3 - Genome
UCSC - Gallus gallus - galGal4 - Genome
UCSC - Gallus gallus - galGal5 - Genome
UCSC - Galeopterus variegatus - balAcu1 - Genome
UCSC - Gasterosteus aculeatus - gasAcu1 - Genome
UCSC - Geospiza fortis - geoFor1 - Genome
UCSC - Gorilla gorilla gorilla - gorGor3 - Genome
UCSC - Gorilla gorilla gorilla - gorGor4 - Genome
UCSC - Gorilla gorilla gorilla - gorGor5 - Genome
UCSC - Homo sapien - hg16 - Genome
UCSC - Homo sapien - hg18 - Genome
UCSC - Homo sapien - hg38 - Genome
UCSC - Homo sapien - GRCh38(hg38) - Genome+RNA
UCSC - Homo sapien - GRCh38(hg38) - RNA
UCSC - Heterocephalus glaber - hetGla1 - Genome
UCSC - Heterocephalus glaber - hetGla2 - Genome
UCSC - Latimeria chalumnae - latCha1 - Genome
UCSC - Loxodonta africana - loxAfr3 - Genome
UCSC - Macaca fascicularis - macFas5 - Genome
UCSC - Manis pentadactyla - manPen1 - Genome
UCSC - Notamacropus eugenii - macEug2 - Genome
UCSC - Melopsittacus undulatus - melUnd1 - Genome
UCSC - Meleagris gallopavo - melGal1 - Genome
UCSC - Meleagris gallopavo - melGal5 - Genome
UCSC - Mustela putorius furo - musFur1 - Genome
UCSC - Microcebus murinus - micMur1 - Genome
UCSC - Microcebus murinus - micMur2 - Genome
UCSC - Mus musculus - mm10 - Genome+RNA
UCSC - Mus musculus - GRCm38(mm10) - RNA
UCSC - Mus musculus - mm10 - Genome
UCSC - Mus musculus - mm7 - Genome
UCSC - Mus musculus - mm8 - Genome
UCSC - Mus musculus - mm9 - Genome
UCSC - Myotis lucifugus - myoLuc2 - Genome
UCSC - Monodelphis domestica - monDom4 - Genome
UCSC - Monodelphis domestica - monDom5 - Genome
UCSC - Nasalis larvatus - nasLar1 - Genome
UCSC - Nanorana parkeri - nanPar1 - Genome
UCSC - Nomascus leucogenys - nomLeu1 - Genome
UCSC - Nomascus leucogenys - nomLeu2 - Genome
UCSC - Nomascus leucogenys - nomLeu3 - Genome
UCSC - Ochotona princeps - ochPri2 - Genome
UCSC - Ochotona princeps - ochPri3 - Genome
UCSC - Otolemur garnettii - otoGar3 - Genome
UCSC - Oryzias latipes - oryLat2 - Genome
UCSC - Oryctolagus cuniculus - oryCun2 - Genome
UCSC - Oreochromis niloticus - oreNil2 - Genome
UCSC - Ornithorhynchus anatinus - ornAna1 - Genome
UCSC - Ornithorhynchus anatinus - ornAna2 - Genome
UCSC - Ovis aries - oviAri1 - Genome
UCSC - Ovis aries - oviAri3 - Genome
UCSC - Pan paniscus - panPan1 - Genome
UCSC - Pan paniscus - panPan2 - Genome
UCSC - Papio anubis - papAnu2 - Genome
UCSC - Papio hamadryas - papHam1 - Genome
UCSC - Pan troglodytes - panTro1 - Genome
UCSC - Pan troglodytes - panTro2 - Genome
UCSC - Pan troglodytes - panTro3 - Genome
UCSC - Pan troglodytes - panTro4 - Genome
UCSC - Pan troglodytes - panTro5 - Genome
UCSC - Petromyzon marinus - petMar1 - Genome
UCSC - Petromyzon marinus - petMar2 - Genome
UCSC - Pteropus vampyrus - pteVam1 - Genome
UCSC - Pongo abelii - ponAbe2 - Genome
UCSC - Pristionchus pacificus (5.0) - priPac1 - Genome
UCSC - Procavia capensis - proCap1 - Genome
UCSC - Rhesus macaque - rheMac2 - Genome
UCSC - Rhesus macaque - rheMac3 - Genome
UCSC - Rhesus macaque - rheMac8 - Genome
UCSC - Rattus norvegicus - rn4 - Genome
UCSC - Rattus norvegicus - rn5 - Genome
UCSC - Rattus norvegicus - rn6 - Genome
UCSC - Rhinopithecus roxellana - rhiRox1 - Genome
UCSC - Saccharomyces cerevisiae - sacCer1 - Genome
UCSC - Saccharomyces cerevisiae - sacCer2 - Genome
UCSC - Saccharomyces cerevisiae - sacCer3 - Genome
UCSC - Sarcophilus harrisii - sarHar1 - Genome
UCSC - Sus scrofa - susScr2 - Genome
UCSC - Sus scrofa - susScr3 - Genome
UCSC - Sus scrofa - susScr11 - Genome
UCSC - Strongylocentrotus purpuratus - strPur1 - Genome
UCSC - Strongylocentrotus purpuratus - strPur2 - Genome
UCSC - Saimiri boliviensis - saiBol1 - Genome
UCSC - Spermophilus tridecemlineatus - speTri2 - Genome
UCSC - Sorex araneus - sorAra1 - Genome
UCSC - Sorex araneus - sorAra2 - Genome
UCSC - Taeniopygia guttata - taeGut1 - Genome
UCSC - Taeniopygia guttata - taeGut2 - Genome
UCSC - Tarsius syrichta - tarSyr1 - Genome
UCSC - Tarsius syrichta - tarSyr2 - Genome
UCSC - Tetraodon nigroviridis - tetNig1 - Genome
UCSC - Tetraodon nigroviridis - tetNig2 - Genome
UCSC - Tupaia belangeri - tupBel1 - Genome
UCSC - Tursiops truncatus - turTru2 - Genome
UCSC - Trichechus manatus latirostris - triMan1 - Genome
UCSC - Vicugna pacos - vicPac1 - Genome
UCSC - Vicugna pacos - vicPac2 - Genome
UCSC - Xenopus laevis - xenLae2 - Genome
UCSC - Xenopus tropicalis - xenTro1 - Genome
UCSC - Xenopus tropicalis - xenTro2 - Genome
UCSC - Xenopus tropicalis - xenTro3 - Genome
UCSC - Xenopus tropicalis - xenTro7 - Genome
UCSC - Xenopus tropicalis - xenTro9 - Genome
Arabidopsis thaliana Genome (TAIR10)
Apis mellifera RNA (honey bee assembly Amel_4.5)
Apis mellifera genome+RNA (honey bee assembly Amel_4.5)
Apis mellifera genome (honey bee assembly Amel_4.5)
Caenorhabditis elegans (assembly WBcel235) RNA
Crassostrea virginica genome (assembly C_virginica-3.0)
Crassostrea virginica RNA (assembly C_virginica-3.0)
Capsicum_annuum (assembly Capsicum_annuum_UCD10X_v.1.0)
C. annuum (assembly Annuum.v1.6.Total)
C. chinense (assembly Chinense.v1.2.Total)
Drosophila melanogaster genome (fruit fly assembly Release 6 plus ISO1 MT)
Danio rerio genome+RNA (zebrafish assembly GRCz11)
Danio rerio genome (zebrafish assembly GRCz11)
Danio rerio RNA (zebrafish assembly GRCz11)
Drosophila melanogaster RNA (fruit fly assembly Release 6 plus ISO1 MT)
Escherichia coli str. K-12 substr. MG1655 (reference genome)
Fragaria vesca v2.0.a1 pseudomolecules
HPV (human papillomavirus reference genomes)
HBV
Klebsiella pneumoniae subsp. pneumoniae HS11286 genome
Mycobacterium tuberculosis H37Rv complete genome
Potato Genome Sequence (DM_v4.04_pseudomolecules)
Pantodon buchholzi mitochondrial DNA (complete genome)
Prunus persica v2.0.a1 scaffolds
Pseudomonas Genomic DNA (pgd_r_16_2)
Phaseolus vulgaris v2.1 genome
Phaseolus vulgaris v2.1 transcript
UNITE Includes singletons set as RefS (version 7.1)
UNITE Includes global and 97% singletons (version 7.1)
Virus genome (EBI)
SILVA-NR99 type strains
Staphylococcus aureus subsp. aureus USA300_FPR3757 ( complete genome)
Sus scrofa (pig) representative genome Sscrofa11.1
Sus scrofa (pig) RNA Sscrofa11.1
Saccharomyces cerevisiae S288C (assembly R64) (reference genome)
Salmonella enterica subsp. enterica serovar Typhi str. CT18 (genome)
Solanum lycopersicum - tomato (assembly SL3.0)
Solanum tuberosum (assembly SolTub_3.0)
Riemerella anatipestifer representative genome
Rattus norvegicus RNA (Norway rat assembly Rnor_6.0)
Rice Genome release 7
C. baccatum(assembly Baccatum.v1.2.Total)
Bubalus bubalis (water buffalo)-All 4 genomes for species
Fragaria_x_ananassa_Genome_v1.0
Sorghum bicolor (assembly Sorghum_bicolor_NCBIv3)
Bovine coronavirus (viruses)
Myxococcus xanthus DK 1622 (complete genome)
Haloferax volcanii (representative genome Haloferax volcanii DS2)
Chlamydia trachomatis (Reference genome Chlamydia trachomatis 434/Bu)
Neisseria gonorrhoeae (Reference genome Neisseria gonorrhoeae FA 1090)
Corynebacterium kutscheri
Streptococcus pneumoniae (Reference genome Streptococcus pneumoniae R60)
Mycoplasma pulmonis (Representative genome Mycoplasma pulmonis UAB CTIP)
Bos_taurus_UMD_3.1.1 assembly for Bos taurus
Salmonella enterica subsp. enterica serovar Typhimurium str. LT2 (genome)
Salmonella enterica subsp. enterica serovar Enteritidis str. P125109 (genome)
Moraxella catarrhalis (All 106 genomes for species)
Spirodela polyrhiza (assembly ASM198140v1)
Bacillus cereus (Reference genome Bacillus cereus ATCC 14579)
Pelodiscus sinensis (assembly PelSin_1.0)
Physcomitrella patens (assembly Phypa V3)
Trichomonas vaginalis G3 (assembly ASM282v1)
Castor canadensis (assembly C.can genome v1.0)
Vaccinia virus WR (complete genome)
African swine fever virus
Clostridium perfringens (Representative genome Clostridium perfringens ATCC 13124)
Stenotrophomonas maltophilia K279a (complete genome)
Zea mays (AGPv4)
longan (Dimocarpus longan Lour.)
Oryza sativa Japonica (IRGSP-1.0)
Oryza sativa Japonica (cDNA)
Leishmania KDNA
Oilseed rape (Brassica napus L.)
Alphavirus (nucleotide)
BosGru_v2.0_genomic
Araport11_genes (cdna)
Medicago truncatula (assembly MedtrA17_4.0)
Oreochromis niloticus (Refseq mRNA)
Manihot esculenta (assembly Manihot esculenta v6)
Moniliophthora perniciosa FA553 (assembly ASM18302v1)
Moniliophthora roreri (assembly ASM146670v1)
Macaca fascicularis (RNA)
Hepacivirus (HCV data set)
Triticum aestivum (bread wheat assembly iwgsc_refseqv1.0)
If we missed databases of your interest, please
tell us
.
Allowed mismatch at the 3' end (exclude the first base)
False
True
Accurate but slow, new in version 3.0.
Output bindingdatas details
False
True
More information about bindingdatas.
T
m
*
Min
°C
Minimum Tm value required for the binding stability between primer and its binding sites.
Size (bp)
From
To
Product size: 0 (min) - 100000 (max).
Concentration of monovalent cations (usually KCl) *
mM
Concentration of divalent cations (usually MgCl
2
) *
mM
Concentration of dNTPs *
mM
Annealing oligo concentration *
nM
Download
MFEprimer-3.0
Specificity
Hairpin
Dimer
Virtual Electrophoretogram
Database Require
Database List