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- 详细信息
- 技术资料
- 保存条件：
-20
- 保质期：
2年
- 英文名：
pRI201-ON
- 库存：
100
- 供应商：
上海烜雅生物科技有限公司
- 规格：
干粉/液体
基本信息
名称：pRI201-ON单子叶植物发根双表达表达质粒
别称: pRI201-ON
质粒属性
质粒简介
pRI201-ON是一个单子叶植物发根农杆菌的双表达质粒。The Ri Plasmid pRI201 is designed for the transformation and expression of target genes from plant cells. This series of binary vectors for plant transformation retains the backbone of the pRI101 vectors, including an alcohol dehydrogenase (ADH) gene-derived 5' untranslated region (5'-UTR) downstream of the cauliflower mosaic virus (CaMV promoter)-derived 35S promoter. Additionally, these vectors have a heat shock protein (HSP) gene-derived terminator in place of the nopaline synthase (NOS) gene-derived terminator, allowing higher target gene expression compared with the pRI101 vector series. Multigene transformation with a single vector can be achieved by integrating an expression cassette containing another gene (promoter + enhancer + gene of interest + terminator) into the vector's second multiple cloning site (MCS2) located downstream of the HSP terminator.
The Ri Plasmid pRI201 binary vector series offers two types of vectors: pRI 201-AN DNA and pRI 201-ON DNA. pRI 201-AN DNA contains an Arabidopsis ADH-derived 5' UTR (AtADH 5'-UTR) and is suitable for dicotyledonous plant transformation. pRI 201-ON DNA contains a rice ADH-derived 5' UTR (OsADH 5'-UTR) and is suitable for monocotyledonous plant transformation. The pRI 201-AN DNA and pRI 201-ON DNA binary vectors for plant transformation have a mutant-type replication origin (Ri ori) from the Rhizobium rhizogenes Ri plasmid. These vectors also contain a replication origin (ColE1 ori) derived from the pUC plasmid; the ColE1 ori allows high copy number replication in E. coli. The vectors' multiple cloning site, located near the right border (RB) of T-DNA, allows stable target gene integration into the plant chromosome.
质粒图谱
质粒序列
LOCUS Exported 10475 bp ds-DNA circular SYN 23-NOV-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..10475
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(57..645)
/direction=LEFT
/label=ori
/note="high-copy-number colE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6419..7213)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin)"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPAT-CPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
promoter 7620..7650
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7658..7674
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 7682..7698
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9075..9091)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
61 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
121 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
181 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
241 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
301 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
361 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
421 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
481 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
541 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
601 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
661 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
721 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
781 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
841 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactcga tcctacaagg
901 tagaatccgc ctgagtcgca agggtgactt cgcctatatt ggacgacggc gcgcagaggg
961 cgacctcttt ttgggttacg attgtaggat tatcactaaa acaatacatg aacatattca
1021 aatggcaatc tctctaaggc attggaaata aatacaaata acagttgggt ggagtttttc
1081 gacctgaggg cgttaacctt ctgttaacct aaaagctctt gcccaaacag cagaatcggc
1141 gctaattgcc agcggcggaa cttttccagt ttcgcgaaaa atatcgccac tggcaaggaa
1201 tgggtttgag atggcgaagt ctgtcctaaa agcagcgcct gtagttgtag ggttgacggc
1261 cttgatggag cgtcatgccg atgccctctc gagccaactt caagcacatc atcttaaggt
1321 tttcccgccg cattccgaga agggtattcg aacattcggg ccatcggagg cgtccaagct
1381 gctcggcgtt ggcgagtcat atttacggca gaccgcgtct gagatgccag agttgaatgt
1441 tagcatgagc ccaggtggca ggcgaatgtt ctcaattgaa gatatccatg tgattcggaa
1501 gtatatggat caggtcggcc gcgggaaccg gcgctacctg ccacatcgtc gaggcggcga
1561 gcagcttcag gttatctctg tgatgaattt caaaggtggg tcgggtaaga ccaccaccgc
1621 cgcgcatctg gcgcagtacc tcgctatgcg cggatatcga gtcttggcca ttgatctcga
1681 tcctcaagcg agcctttctg cactctttgg gagccaaccg gagacggacg ttggcccgaa
1741 cgaaacgctc tacggcgcta taaggtatga tgatgagcag gtggcaatcg aacgagtcgt
1801 ccgagggact tacattcccg acctccacct gattcctggt aaccttgagc tgatggagtt
1861 tgaacacgat acgccacgcg cgctgatgaa ccgcaaagag ggcgacacgc tcttttatgg
1921 tcgcatcagc caagtaattg aagatatcgc ggataactat gacgtcgtgg tcatcgactg
1981 ccctccccag cttgggtatc tcacgctatc cgcattgact gcggcgacgt ccattcttgt
2041 cacggtccat ccgcagatgc tggatgtgat gtcgatgaac cagtttctgg caatgacatc
2101 gaaccttttg cgtgaaatcg agaatgctgg cgccaagttc aagtttaatt ggatgcgcta
2161 tctgataacc cgtttcgaac cgagcgacgg accacagaac caaatggtag gttatctgcg
2221 gtcgattttt ggcgaaaatg tcctcaattt tccgatgctt aaaaccaccg cggtttcgga
2281 cgctggcctg acaaaccaga ctctattcga agtggagcgt ggcctgttca cgcgctcgac
2341 ctatgatcga gccttggagg cgatgaacgc cgtcaacgac gagatcgaaa cactgatcaa
2401 aaaagcatgg ggtaggccca catgagccgg aagcacatcc ttggcgtctc aactgacgcc
2461 cctgagacgt cgcccgccga caataggacg gcaaagaacc gctccatgcc gctcctcggc
2521 gtaacaagga aggagcgcga tccggcaacg aagctcacag cgaacattgg taacgcactg
2581 cgagagcaaa acgatcgtct tagccgtgcc gaagagatcg agcggcgtct cgctgaaggt
2641 caggcagtga tagagttgga tgcctcgtca atagaaccgt ctttcgtgca ggatcgtatg
2701 cgaggggaca ttgacgggct ccttacttcg atccgggaac aaggacagca agtcccaatc
2761 cttgtgcgac cgcatccgag ccagccgggc cgatatcagg ttgccttcgg ccaccgccgg
2821 ctacgcgccg tttcagaact cggacttccg gtcagggcgg tcgttcgcga actgacggac
2881 gagcaagtgg tcgtagcaca gggtcaggaa aacaatgtgc gcgaagatct taccttcatc
2941 gaaaaggcgc gcttcgcaca tcgcctgaac aggcagtttt ctcgagagat tgtcatcgcc
3001 gcgatgtcga tcgacaagag caatttgtcc aagatgcttc tgctcgttga cgccctcccc
3061 tctgaactga ccgatgctat tggtgccgct cctggtgttg gacggccgag ttggcaacaa
3121 cttgccgagc tgattgagaa agtttcttca ccggccgacg tggctaaata tgctatgtcg
3181 gaggaagttc aagcgctgcc atcggcagaa cgattcaagg cggtgatcgc tagtctgaag
3241 cccagtcggg ttgcgcgtgg acttcccgag gtcatggcca ccccagacgg caccagaatt
3301 gcacaggtga cgcagagcaa ggccaaactg gaaatcacga ttgacaggaa ggcgacgccc
3361 gattttgcga ccttcgtgct cgatcatgtg ccagcgctgt atcaagcgta ccacgctgag
3421 aaccaacgga aacggggaga gtaaaccgca aaagaaaaga gccccctcaa cgtcgccgtc
3481 gcggaagccc ttctgtctct ctagcgcgaa cagaatcgca tttcctcgaa tcctcgtcaa
3541 gagtttttag cgccgttttg gtgagctgat ttcctttgcc tgctgaaagg tgaaagatga
3601 tgcagacagg aagtgtaacg acgccattcg ggcggcggcc aatgacgctt gcgcttgtgc
3661 ggcgccagac ggcgctggcc gatatcaaac aaggcaagac agcggacaag tggaaggtct
3721 ttagggacgc gtccgcggcc atggaactac ttggaatcca gtccaacagt cttgccgtcc
3781 ttgatgcgct attgagcttt cacccggaaa cggagttgcg tcaggaggca cagctgatcg
3841 tcttcccgtc gaatgctcag cttgcccttc gggcgcatgg gatggctggc gcgactttgc
3901 gtaggcacat cgccatgctc gtggagtcag gcttgatcgt ccggaaggat agcgccaacg
3961 gaaagcgtta cgctcgtaag gatggcgctg gtcagatcga gcgcgcgttt ggcttcgatt
4021 tgtctccgct tctcgcgcgg tccgaagagc tagcgatgat ggcacagcag gtgatggccg
4081 atcgagcagc attcaggatg gccaaagaaa gtctgacgat ttgccgacgg gacgttcgga
4141 agctaattac ggcagctatg gaagagggag cggagggcga ctggcaagct gtcgaggaag
4201 tctatgtgga acttgtgggt agaatcccac gcgccccgac gcttgctgat gtagagtcaa
4261 ttctcgaaga gatgtggatg ctccaggaag agataatcaa ccggttggaa attagagaca
4321 attcagaaaa taatagcacc aatgctgccc agagcgagca gcacatacag aactcaaaac
4381 ccgaatccgt taatgaactt gaacctcgct ctgaaaagga gcagggcgct aagccgagtg
4441 aaatagaccg ggcaaggagc gagccgataa aagcgttccc cctcgggatg atcctgaaag
4501 cgtgcccgac cattggcaat tatgggccga gcggtgcggt tgctagctgg cgtgacctca
4561 tgtcggctgc ggtggtggtt cggtctatgc tgggggtcag cccgtcggct taccaagacg
4621 cgtgtgaggc aatgggaccg gagaatgcgg cagcagcgat ggcgtgcatt ttggagcgag
4681 cgaacttcat caattcgcct gggggctatc tccgagatct gacacggcgg agcgaactcg
4741 ggaagttttc acttggcccg atgataatgg cgctcttgaa ggctagcggg caggggacgt
4801 tgcggtttgg ctagaattag cgagtatgga gcaggatggt ctgtggtcag ctgaccacag
4861 acctaatagg ttgaaaacat gagcgttttt tggatgatcg acagaccatc cgattcccgg
4921 agtaccaagc gtgctctgat gggagcgata acattactca acaagcacga aggccccatg
4981 ccgatcgttg atcgtgaagg agagcctgct ctacatgcgg cggtattttg ccggccgagg
5041 catgtagtcg cggagcactg cctatttact gccctaggca caaacgttga ctcttggatc
5101 gagctggcag acaaagcaat aacccacaca gaggacgatt aatggctgac gaagagatcc
5161 agaatccgcc ggacggtact gctgctgccg aagttgagcc ggctgctcct agaggtagaa
5221 gagcaaagaa agcaccagcc gaaacagccc gcacgggatc gttcaaatcc gtgaagccga
5281 aaacccgcgg cctcagcaac cgagaaaaac tggagaagat cggtcaaatc gaagctcagg
5341 tcgctggcgg cgcaaccttg aaggacgccg ttaagatcgt gggtatttcc gttcagacct
5401 attatcaatg gaagagagct gcggttcaac ctgtctcaca gaatccggcc gtgtctgttt
5461 cagttgacga tgaactcggc gagttcatcc aactcgagga ggaaaatcgg cggctcagaa
5521 agctagcttg tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata
5581 acacattgcg gacgttttta atgtactggg gtggtttttc ttttcaccag tgagacgggc
5641 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
5701 gtttgcccca gcaggcgaaa atcctgtttg atggtggttc cgaaatcggc aaaatccctt
5761 ataaatcaaa agaatagccc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
5821 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
5881 gcccaaattc ccgatctagt aacatagatg acaccgcgcg cgataattta tcctagtttg
5941 cgcgctatat tttgttttct atcgcgtatt aaatgtataa ttgcgggact ctaatcataa
6001 aaacccatct cataaataac gtcatgcatt acatgttaat tattacatgc ttaacgtaat
6061 tcaacagaaa ttatatgata atcatcgcaa gaccggcaac aggattcaat cttaagaaac
6121 tttattgcca aatgtttgaa cgatcggggc ccgatcatat tgtcgctcag gatcgtggcg
6181 ttgtgcttgt cggccgttgc tgtcgtaatg atatcggcac cttcgaccgc ctgttccgca
6241 gagatcccgt gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg
6301 gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa
6361 tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc
6421 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
6481 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg
6541 tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc
6601 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga
6661 cgagatcatc gccgtcgggc atacgcgcct tgagcctggc gaacagttcg gctggcgcga
6721 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
6781 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
6841 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag
6901 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
6961 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
7021 ctgcctcgtc ctgtagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
7081 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
7141 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat
7201 cttgttcaat catgcgaaac gatccagatc cggtgcagat tatttggatt gagagtgaat
7261 atgagactct aattggatac cgaggggaat ttatggaacg tcagtggagc atttttgaca
7321 agaaatattt gctagctgat agtgacctta ggcgactttt gaacgcgcaa taatggtttc
7381 tgacgtatgt gcttagctca ttaaactcca gaaacccgcg gctgagtggc tccttcaacg
7441 ttgcggttct gtcagttcca aacgtaaaac ggcttgtccc gcgtcatcgg cgggggtcat
7501 aacgtgactc ccttaattct ccgctcatga tcctggcacg acaggtttcc cgactggaaa
7561 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
7621 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
7681 acaggaaaca gctatgacca tgattacgaa ttcccgggat ccagcggccg caggtaccaa
7741 ttccttatct ttaatcatat tccatagtcc ataccatagc acatacagta gttatatgcg
7801 aagagatcca acaaaacatt cacaatggat tatagaaaca tttgtttatt cattataatg
7861 agatcttaca ttcatttaat attagaaaaa gccacaaatt cataacacaa caagccaaga
7921 aaaaaacaca aacttaagca cacaagctag ctttttattt gacacaccaa atatttcatc
7981 ttcatcttca taagagctcg gtagcaattc ccgaggctgt agccgacgat ggtgcgccag
8041 gagagttgtt gattcagaat tgtcgacccg gtagtaactt gctaatcgac gggcatatgt
8101 aatccccctc tttttcaaag aacaagaaac cactccgtag agatcgaaag ctcaggtttt
8161 cttttttctt gaatcactcg cagttcgttg cttggaattc tctagagtcc cccgtgttct
8221 ctccaaatga aatgaacttc cttatataga ggaagggtct tgcgaaggat agtgggattg
8281 tgcgtcatcc cttacgtcag tggagatatc acatcaatcc acttgctttg aagacgtggt
8341 tggaacgtct tctttttcca cgatgctcct cgtgggtggg ggtccatctt tgggaccact
8401 gtcggcagag gcatcttcaa cgatggcctt tcctttatcg caatgatggc atttgtagga
8461 gccaccttcc ttttccacta tcttcacaat aaagtgacag atagctgggc aatggaatcc
8521 gaggaggttt ccggatatta ccctttgttg aaaagtctca attgcccttt ggtcttctga
8581 gactgtatct ttgatatttt tggagtagac aagtgtgtcg tgctccacca tgttgacgaa
8641 gattttcttc ttgtcattga gtcgtaagag actctgtatg aactgttcgc cagtctttac
8701 ggcgagttct gttaggtcct ctatttgaat ctttgactcc atggcctttg attcagtggg
8761 aactaccttt ttagagactc caatctctat tacttgcctt ggtttgtgaa gcaagccttg
8821 aatcgtccat actggaatag tacttctgat cttgagaaat atatctttct ctgtgttctt
8881 gatgcagtta gtcctgaatc ttttgactgc atctttaacc ttcttgggaa ggtatttgat
8941 ttcctggaga ttattgctcg ggtagatcgt cttgatgaga cctgctgcgt aagcctctct
9001 aaccatctgt gggttagcat tctttctgaa attgaaaagg ctaatctggg gacctgcagg
9061 catgcaagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
9121 acccaactta atcgccttgc agcacatccc cctttcgcca ggatcagatt gtcgtttccc
9181 gccttcagtt taaactatca gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa
9241 agagcgttta ttagaataat cggatattta aaagggcgtg aaaaggttta tccgttcgtc
9301 catttgtatg tgactaaaac aattcatcca gtaaaatata atattttatt ttctcccaat
9361 caggcttgat ccccagtaag tcaaaaaata gctcgacata ctgttcttcc ccgatatcct
9421 ccctgatcga ccggacgcag aaggcaatgt cataccactt gtccgccctg ccgcttctcc
9481 caagatcaat aaagccactt actttgccat ctttcacaaa gatgttgctg tctcccaggt
9541 cgccgtggga aaagacaagt tcctcttcgg gcttttccgt ctttaaaaaa tcatacagct
9601 cgcgcggatc tttaaatgga gtgtcttctt cccagttttc gcaatccaca tcggccagat
9661 cgttattcag taagtaatcc aattcggcta agcggctgtc taagctattc gtatagggac
9721 aatccgatat gtcgatggag tgaaagagcc tgatgcactc cgcatacagc tcgataatct
9781 tttcagggct ttgttcatct tcatactctt ccgagcaaag gacgccatcg gcctcactca
9841 tgagcagatt gctccagcca tcatgccgtt caaagtgcag gacctttgga acaggcagct
9901 ttccttccag ccatagcatc atgtcctttt cccgttccac atcataggtg gtccctttat
9961 accggctgtc cgtcattttt aaatataggt tttcattttc tcccaccagc ttatatacct
10021 tagcaggaga cattccttcc gtatctttta cgcagcggta tttttcgatc agttttttca
10081 attccggtga tattctcatt ttagccattt attatttcct tcctcttttc tacagtattt
10141 aaagataccc caagaagcta attataacaa gacgaactcc aattcactgt tccttgcatt
10201 ctaaaacctt aaataccaga aaacagcttt ttcaaagttg ttttcaaagt tggcgtataa
10261 catagtatcg acggagccga ttttgaaacc acaattatgg gtgatgctgc caacttactg
10321 atttagtgta tgatggtgtt tttgaggtgc tccagtggct tctgtttcta tcagctgtcc
10381 ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg gacatcagcg
10441 ctatctctgc tctcactgcc gtaaaacatg gcaac
//
质粒菌株产品操作说明书
一、扩增流程
收到产品后，请先根据产品管壁标签来判断产品形式，并在扩增前准确查找该质粒菌株的抗性、感受态和培养温度。
1、质粒干粉（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
①收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O去离子水溶解质粒；
②取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；（从第二步开始均要在超净工作台中无菌操作）
③加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min；
④6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上；（可使用本平台的平板涂布专用玻璃珠进行涂布，可以比传统涂布方法获得更多转化子）
⑤将平板正向培养1h，再倒置37℃培养14h。如果要求是30度则培养20h；
（菌落过多则将质粒稀释后再转化。没有菌落则加入10μl质粒转化。另不要直接转表达感受态，要先转克隆感受态，重提质粒后再导入表达感受态）
⑥挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
2、甘油菌种（冰袋运输，存于-80℃，保质期90天，请务必划线挑单克隆培养）
四区划线后挑单菌落培养，酵母菌需要先液体复苏再四区划线，再挑单菌落液体培养。
3、穿刺菌种（冰袋运输，存于4℃，保质期7天）
穿刺接种，液体培养后四区划线，再挑单菌落液体培养。
4、菌落平板（冰袋运输，存于4℃，保质期7天）
直接挑取单菌落至液体培养基中。
5、液体质粒（冰袋运输，存于-20℃，保质期90天）
单独提取的液体质粒收到后可直接使用。
6、滤纸质粒（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
收到货后将滤纸画圈部分剪下放入EP管中，加100ul无菌水将滤纸浸湿并浸泡5min，吸取5ul质粒转化，离心全涂。
二、转化图片
名称：pRI201-ON单子叶植物发根双表达表达质粒
别称: pRI201-ON
| 启动子: | 35s |
|---|---|
| 复制子: | RI |
| 原核抗性: | Kan |
| 筛选标记: | G418 |
| 克隆菌株: | Stbl3 |
| 培养条件: | 37度 |
质粒属性
| 载体宿主： | 植物 |
|---|---|
| 载体用途： | 蛋白表达 |
| 基因种属： | |
| 基因类型： | ORF |
| 原核抗性： | Kan |
| 筛选标记： | Neo/G418 |
| 荧光蛋白： |
质粒简介
pRI201-ON是一个单子叶植物发根农杆菌的双表达质粒。The Ri Plasmid pRI201 is designed for the transformation and expression of target genes from plant cells. This series of binary vectors for plant transformation retains the backbone of the pRI101 vectors, including an alcohol dehydrogenase (ADH) gene-derived 5' untranslated region (5'-UTR) downstream of the cauliflower mosaic virus (CaMV promoter)-derived 35S promoter. Additionally, these vectors have a heat shock protein (HSP) gene-derived terminator in place of the nopaline synthase (NOS) gene-derived terminator, allowing higher target gene expression compared with the pRI101 vector series. Multigene transformation with a single vector can be achieved by integrating an expression cassette containing another gene (promoter + enhancer + gene of interest + terminator) into the vector's second multiple cloning site (MCS2) located downstream of the HSP terminator.
The Ri Plasmid pRI201 binary vector series offers two types of vectors: pRI 201-AN DNA and pRI 201-ON DNA. pRI 201-AN DNA contains an Arabidopsis ADH-derived 5' UTR (AtADH 5'-UTR) and is suitable for dicotyledonous plant transformation. pRI 201-ON DNA contains a rice ADH-derived 5' UTR (OsADH 5'-UTR) and is suitable for monocotyledonous plant transformation. The pRI 201-AN DNA and pRI 201-ON DNA binary vectors for plant transformation have a mutant-type replication origin (Ri ori) from the Rhizobium rhizogenes Ri plasmid. These vectors also contain a replication origin (ColE1 ori) derived from the pUC plasmid; the ColE1 ori allows high copy number replication in E. coli. The vectors' multiple cloning site, located near the right border (RB) of T-DNA, allows stable target gene integration into the plant chromosome.
质粒图谱
质粒序列
LOCUS Exported 10475 bp ds-DNA circular SYN 23-NOV-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..10475
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(57..645)
/direction=LEFT
/label=ori
/note="high-copy-number colE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6419..7213)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin)"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPAT-CPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
promoter 7620..7650
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7658..7674
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 7682..7698
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9075..9091)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
61 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
121 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
181 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
241 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
301 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
361 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
421 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
481 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
541 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
601 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
661 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
721 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
781 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
841 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactcga tcctacaagg
901 tagaatccgc ctgagtcgca agggtgactt cgcctatatt ggacgacggc gcgcagaggg
961 cgacctcttt ttgggttacg attgtaggat tatcactaaa acaatacatg aacatattca
1021 aatggcaatc tctctaaggc attggaaata aatacaaata acagttgggt ggagtttttc
1081 gacctgaggg cgttaacctt ctgttaacct aaaagctctt gcccaaacag cagaatcggc
1141 gctaattgcc agcggcggaa cttttccagt ttcgcgaaaa atatcgccac tggcaaggaa
1201 tgggtttgag atggcgaagt ctgtcctaaa agcagcgcct gtagttgtag ggttgacggc
1261 cttgatggag cgtcatgccg atgccctctc gagccaactt caagcacatc atcttaaggt
1321 tttcccgccg cattccgaga agggtattcg aacattcggg ccatcggagg cgtccaagct
1381 gctcggcgtt ggcgagtcat atttacggca gaccgcgtct gagatgccag agttgaatgt
1441 tagcatgagc ccaggtggca ggcgaatgtt ctcaattgaa gatatccatg tgattcggaa
1501 gtatatggat caggtcggcc gcgggaaccg gcgctacctg ccacatcgtc gaggcggcga
1561 gcagcttcag gttatctctg tgatgaattt caaaggtggg tcgggtaaga ccaccaccgc
1621 cgcgcatctg gcgcagtacc tcgctatgcg cggatatcga gtcttggcca ttgatctcga
1681 tcctcaagcg agcctttctg cactctttgg gagccaaccg gagacggacg ttggcccgaa
1741 cgaaacgctc tacggcgcta taaggtatga tgatgagcag gtggcaatcg aacgagtcgt
1801 ccgagggact tacattcccg acctccacct gattcctggt aaccttgagc tgatggagtt
1861 tgaacacgat acgccacgcg cgctgatgaa ccgcaaagag ggcgacacgc tcttttatgg
1921 tcgcatcagc caagtaattg aagatatcgc ggataactat gacgtcgtgg tcatcgactg
1981 ccctccccag cttgggtatc tcacgctatc cgcattgact gcggcgacgt ccattcttgt
2041 cacggtccat ccgcagatgc tggatgtgat gtcgatgaac cagtttctgg caatgacatc
2101 gaaccttttg cgtgaaatcg agaatgctgg cgccaagttc aagtttaatt ggatgcgcta
2161 tctgataacc cgtttcgaac cgagcgacgg accacagaac caaatggtag gttatctgcg
2221 gtcgattttt ggcgaaaatg tcctcaattt tccgatgctt aaaaccaccg cggtttcgga
2281 cgctggcctg acaaaccaga ctctattcga agtggagcgt ggcctgttca cgcgctcgac
2341 ctatgatcga gccttggagg cgatgaacgc cgtcaacgac gagatcgaaa cactgatcaa
2401 aaaagcatgg ggtaggccca catgagccgg aagcacatcc ttggcgtctc aactgacgcc
2461 cctgagacgt cgcccgccga caataggacg gcaaagaacc gctccatgcc gctcctcggc
2521 gtaacaagga aggagcgcga tccggcaacg aagctcacag cgaacattgg taacgcactg
2581 cgagagcaaa acgatcgtct tagccgtgcc gaagagatcg agcggcgtct cgctgaaggt
2641 caggcagtga tagagttgga tgcctcgtca atagaaccgt ctttcgtgca ggatcgtatg
2701 cgaggggaca ttgacgggct ccttacttcg atccgggaac aaggacagca agtcccaatc
2761 cttgtgcgac cgcatccgag ccagccgggc cgatatcagg ttgccttcgg ccaccgccgg
2821 ctacgcgccg tttcagaact cggacttccg gtcagggcgg tcgttcgcga actgacggac
2881 gagcaagtgg tcgtagcaca gggtcaggaa aacaatgtgc gcgaagatct taccttcatc
2941 gaaaaggcgc gcttcgcaca tcgcctgaac aggcagtttt ctcgagagat tgtcatcgcc
3001 gcgatgtcga tcgacaagag caatttgtcc aagatgcttc tgctcgttga cgccctcccc
3061 tctgaactga ccgatgctat tggtgccgct cctggtgttg gacggccgag ttggcaacaa
3121 cttgccgagc tgattgagaa agtttcttca ccggccgacg tggctaaata tgctatgtcg
3181 gaggaagttc aagcgctgcc atcggcagaa cgattcaagg cggtgatcgc tagtctgaag
3241 cccagtcggg ttgcgcgtgg acttcccgag gtcatggcca ccccagacgg caccagaatt
3301 gcacaggtga cgcagagcaa ggccaaactg gaaatcacga ttgacaggaa ggcgacgccc
3361 gattttgcga ccttcgtgct cgatcatgtg ccagcgctgt atcaagcgta ccacgctgag
3421 aaccaacgga aacggggaga gtaaaccgca aaagaaaaga gccccctcaa cgtcgccgtc
3481 gcggaagccc ttctgtctct ctagcgcgaa cagaatcgca tttcctcgaa tcctcgtcaa
3541 gagtttttag cgccgttttg gtgagctgat ttcctttgcc tgctgaaagg tgaaagatga
3601 tgcagacagg aagtgtaacg acgccattcg ggcggcggcc aatgacgctt gcgcttgtgc
3661 ggcgccagac ggcgctggcc gatatcaaac aaggcaagac agcggacaag tggaaggtct
3721 ttagggacgc gtccgcggcc atggaactac ttggaatcca gtccaacagt cttgccgtcc
3781 ttgatgcgct attgagcttt cacccggaaa cggagttgcg tcaggaggca cagctgatcg
3841 tcttcccgtc gaatgctcag cttgcccttc gggcgcatgg gatggctggc gcgactttgc
3901 gtaggcacat cgccatgctc gtggagtcag gcttgatcgt ccggaaggat agcgccaacg
3961 gaaagcgtta cgctcgtaag gatggcgctg gtcagatcga gcgcgcgttt ggcttcgatt
4021 tgtctccgct tctcgcgcgg tccgaagagc tagcgatgat ggcacagcag gtgatggccg
4081 atcgagcagc attcaggatg gccaaagaaa gtctgacgat ttgccgacgg gacgttcgga
4141 agctaattac ggcagctatg gaagagggag cggagggcga ctggcaagct gtcgaggaag
4201 tctatgtgga acttgtgggt agaatcccac gcgccccgac gcttgctgat gtagagtcaa
4261 ttctcgaaga gatgtggatg ctccaggaag agataatcaa ccggttggaa attagagaca
4321 attcagaaaa taatagcacc aatgctgccc agagcgagca gcacatacag aactcaaaac
4381 ccgaatccgt taatgaactt gaacctcgct ctgaaaagga gcagggcgct aagccgagtg
4441 aaatagaccg ggcaaggagc gagccgataa aagcgttccc cctcgggatg atcctgaaag
4501 cgtgcccgac cattggcaat tatgggccga gcggtgcggt tgctagctgg cgtgacctca
4561 tgtcggctgc ggtggtggtt cggtctatgc tgggggtcag cccgtcggct taccaagacg
4621 cgtgtgaggc aatgggaccg gagaatgcgg cagcagcgat ggcgtgcatt ttggagcgag
4681 cgaacttcat caattcgcct gggggctatc tccgagatct gacacggcgg agcgaactcg
4741 ggaagttttc acttggcccg atgataatgg cgctcttgaa ggctagcggg caggggacgt
4801 tgcggtttgg ctagaattag cgagtatgga gcaggatggt ctgtggtcag ctgaccacag
4861 acctaatagg ttgaaaacat gagcgttttt tggatgatcg acagaccatc cgattcccgg
4921 agtaccaagc gtgctctgat gggagcgata acattactca acaagcacga aggccccatg
4981 ccgatcgttg atcgtgaagg agagcctgct ctacatgcgg cggtattttg ccggccgagg
5041 catgtagtcg cggagcactg cctatttact gccctaggca caaacgttga ctcttggatc
5101 gagctggcag acaaagcaat aacccacaca gaggacgatt aatggctgac gaagagatcc
5161 agaatccgcc ggacggtact gctgctgccg aagttgagcc ggctgctcct agaggtagaa
5221 gagcaaagaa agcaccagcc gaaacagccc gcacgggatc gttcaaatcc gtgaagccga
5281 aaacccgcgg cctcagcaac cgagaaaaac tggagaagat cggtcaaatc gaagctcagg
5341 tcgctggcgg cgcaaccttg aaggacgccg ttaagatcgt gggtatttcc gttcagacct
5401 attatcaatg gaagagagct gcggttcaac ctgtctcaca gaatccggcc gtgtctgttt
5461 cagttgacga tgaactcggc gagttcatcc aactcgagga ggaaaatcgg cggctcagaa
5521 agctagcttg tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata
5581 acacattgcg gacgttttta atgtactggg gtggtttttc ttttcaccag tgagacgggc
5641 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
5701 gtttgcccca gcaggcgaaa atcctgtttg atggtggttc cgaaatcggc aaaatccctt
5761 ataaatcaaa agaatagccc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
5821 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
5881 gcccaaattc ccgatctagt aacatagatg acaccgcgcg cgataattta tcctagtttg
5941 cgcgctatat tttgttttct atcgcgtatt aaatgtataa ttgcgggact ctaatcataa
6001 aaacccatct cataaataac gtcatgcatt acatgttaat tattacatgc ttaacgtaat
6061 tcaacagaaa ttatatgata atcatcgcaa gaccggcaac aggattcaat cttaagaaac
6121 tttattgcca aatgtttgaa cgatcggggc ccgatcatat tgtcgctcag gatcgtggcg
6181 ttgtgcttgt cggccgttgc tgtcgtaatg atatcggcac cttcgaccgc ctgttccgca
6241 gagatcccgt gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg
6301 gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa
6361 tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc
6421 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
6481 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg
6541 tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc
6601 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga
6661 cgagatcatc gccgtcgggc atacgcgcct tgagcctggc gaacagttcg gctggcgcga
6721 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
6781 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
6841 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag
6901 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
6961 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
7021 ctgcctcgtc ctgtagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
7081 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
7141 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat
7201 cttgttcaat catgcgaaac gatccagatc cggtgcagat tatttggatt gagagtgaat
7261 atgagactct aattggatac cgaggggaat ttatggaacg tcagtggagc atttttgaca
7321 agaaatattt gctagctgat agtgacctta ggcgactttt gaacgcgcaa taatggtttc
7381 tgacgtatgt gcttagctca ttaaactcca gaaacccgcg gctgagtggc tccttcaacg
7441 ttgcggttct gtcagttcca aacgtaaaac ggcttgtccc gcgtcatcgg cgggggtcat
7501 aacgtgactc ccttaattct ccgctcatga tcctggcacg acaggtttcc cgactggaaa
7561 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
7621 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
7681 acaggaaaca gctatgacca tgattacgaa ttcccgggat ccagcggccg caggtaccaa
7741 ttccttatct ttaatcatat tccatagtcc ataccatagc acatacagta gttatatgcg
7801 aagagatcca acaaaacatt cacaatggat tatagaaaca tttgtttatt cattataatg
7861 agatcttaca ttcatttaat attagaaaaa gccacaaatt cataacacaa caagccaaga
7921 aaaaaacaca aacttaagca cacaagctag ctttttattt gacacaccaa atatttcatc
7981 ttcatcttca taagagctcg gtagcaattc ccgaggctgt agccgacgat ggtgcgccag
8041 gagagttgtt gattcagaat tgtcgacccg gtagtaactt gctaatcgac gggcatatgt
8101 aatccccctc tttttcaaag aacaagaaac cactccgtag agatcgaaag ctcaggtttt
8161 cttttttctt gaatcactcg cagttcgttg cttggaattc tctagagtcc cccgtgttct
8221 ctccaaatga aatgaacttc cttatataga ggaagggtct tgcgaaggat agtgggattg
8281 tgcgtcatcc cttacgtcag tggagatatc acatcaatcc acttgctttg aagacgtggt
8341 tggaacgtct tctttttcca cgatgctcct cgtgggtggg ggtccatctt tgggaccact
8401 gtcggcagag gcatcttcaa cgatggcctt tcctttatcg caatgatggc atttgtagga
8461 gccaccttcc ttttccacta tcttcacaat aaagtgacag atagctgggc aatggaatcc
8521 gaggaggttt ccggatatta ccctttgttg aaaagtctca attgcccttt ggtcttctga
8581 gactgtatct ttgatatttt tggagtagac aagtgtgtcg tgctccacca tgttgacgaa
8641 gattttcttc ttgtcattga gtcgtaagag actctgtatg aactgttcgc cagtctttac
8701 ggcgagttct gttaggtcct ctatttgaat ctttgactcc atggcctttg attcagtggg
8761 aactaccttt ttagagactc caatctctat tacttgcctt ggtttgtgaa gcaagccttg
8821 aatcgtccat actggaatag tacttctgat cttgagaaat atatctttct ctgtgttctt
8881 gatgcagtta gtcctgaatc ttttgactgc atctttaacc ttcttgggaa ggtatttgat
8941 ttcctggaga ttattgctcg ggtagatcgt cttgatgaga cctgctgcgt aagcctctct
9001 aaccatctgt gggttagcat tctttctgaa attgaaaagg ctaatctggg gacctgcagg
9061 catgcaagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
9121 acccaactta atcgccttgc agcacatccc cctttcgcca ggatcagatt gtcgtttccc
9181 gccttcagtt taaactatca gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa
9241 agagcgttta ttagaataat cggatattta aaagggcgtg aaaaggttta tccgttcgtc
9301 catttgtatg tgactaaaac aattcatcca gtaaaatata atattttatt ttctcccaat
9361 caggcttgat ccccagtaag tcaaaaaata gctcgacata ctgttcttcc ccgatatcct
9421 ccctgatcga ccggacgcag aaggcaatgt cataccactt gtccgccctg ccgcttctcc
9481 caagatcaat aaagccactt actttgccat ctttcacaaa gatgttgctg tctcccaggt
9541 cgccgtggga aaagacaagt tcctcttcgg gcttttccgt ctttaaaaaa tcatacagct
9601 cgcgcggatc tttaaatgga gtgtcttctt cccagttttc gcaatccaca tcggccagat
9661 cgttattcag taagtaatcc aattcggcta agcggctgtc taagctattc gtatagggac
9721 aatccgatat gtcgatggag tgaaagagcc tgatgcactc cgcatacagc tcgataatct
9781 tttcagggct ttgttcatct tcatactctt ccgagcaaag gacgccatcg gcctcactca
9841 tgagcagatt gctccagcca tcatgccgtt caaagtgcag gacctttgga acaggcagct
9901 ttccttccag ccatagcatc atgtcctttt cccgttccac atcataggtg gtccctttat
9961 accggctgtc cgtcattttt aaatataggt tttcattttc tcccaccagc ttatatacct
10021 tagcaggaga cattccttcc gtatctttta cgcagcggta tttttcgatc agttttttca
10081 attccggtga tattctcatt ttagccattt attatttcct tcctcttttc tacagtattt
10141 aaagataccc caagaagcta attataacaa gacgaactcc aattcactgt tccttgcatt
10201 ctaaaacctt aaataccaga aaacagcttt ttcaaagttg ttttcaaagt tggcgtataa
10261 catagtatcg acggagccga ttttgaaacc acaattatgg gtgatgctgc caacttactg
10321 atttagtgta tgatggtgtt tttgaggtgc tccagtggct tctgtttcta tcagctgtcc
10381 ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg gacatcagcg
10441 ctatctctgc tctcactgcc gtaaaacatg gcaac
//
质粒菌株产品操作说明书
一、扩增流程
收到产品后，请先根据产品管壁标签来判断产品形式，并在扩增前准确查找该质粒菌株的抗性、感受态和培养温度。
1、质粒干粉（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
①收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O去离子水溶解质粒；
②取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；（从第二步开始均要在超净工作台中无菌操作）
③加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min；
④6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上；（可使用本平台的平板涂布专用玻璃珠进行涂布，可以比传统涂布方法获得更多转化子）
⑤将平板正向培养1h，再倒置37℃培养14h。如果要求是30度则培养20h；
（菌落过多则将质粒稀释后再转化。没有菌落则加入10μl质粒转化。另不要直接转表达感受态，要先转克隆感受态，重提质粒后再导入表达感受态）
⑥挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
2、甘油菌种（冰袋运输，存于-80℃，保质期90天，请务必划线挑单克隆培养）
四区划线后挑单菌落培养，酵母菌需要先液体复苏再四区划线，再挑单菌落液体培养。
3、穿刺菌种（冰袋运输，存于4℃，保质期7天）
穿刺接种，液体培养后四区划线，再挑单菌落液体培养。
4、菌落平板（冰袋运输，存于4℃，保质期7天）
直接挑取单菌落至液体培养基中。
5、液体质粒（冰袋运输，存于-20℃，保质期90天）
单独提取的液体质粒收到后可直接使用。
6、滤纸质粒（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
收到货后将滤纸画圈部分剪下放入EP管中，加100ul无菌水将滤纸浸湿并浸泡5min，吸取5ul质粒转化，离心全涂。
二、转化图片
| P0043/pET-50b大肠表达质粒 |
| P0996/pET-51b大肠表达质粒 |
| P0044/pET-52b大肠表达质粒 |
| P0045/pET302/NT-His大肠表达质粒 |
| P1734/pET303/CT-His大肠表达质粒 |
| P0046/pETM-11大肠表达质粒 |
| P0047/pETM-30大肠表达质粒 |
| P0998/pETM-41大肠表达质粒 |
| P0336/pETBlue-2大肠表达质粒 |
| P1678/pETcoco-1大肠表达质粒 |
风险提示：丁香通仅作为第三方平台，为商家信息发布提供平台空间。用户咨询产品时请注意保护个人信息及财产安全，合理判断，谨慎选购商品，商家和用户对交易行为负责。对于医疗器械类产品，请先查证核实企业经营资质和医疗器械产品注册证情况。
pRI201-ON单子叶植物发根双表达表达质粒
￥100 - 1000
