pCMV-Scn7a-3×FLAG-Neo plasmid

PVT97516

pCMV-Scn7a-3×FLAG-Neo plasmid Information

Plasmid name: pCMV-Scn7a(mouse)-3×FLAG-Neo
Alias: NM_009135.2;1110034K09Rik; NaG; Nav2; Nav2.3; Nax;Scn6a;
Gene lenght: 5043bp
Host: Mammalian cells
Use(s): Protein Expression
Fragment type: ORF
Species: Mouse
Prokaryotic resistance: Amp
Screening marker: Neo/G418
Fluorescent labeling:
Promoter: CMV
Replicon: pUC
Growth Strain(s): BL21(DE3)
Growth Temperature: 30°C
Vector backbone:
Forward primer: CMV-F
Reverse primer:
Copy number: Low
Induction:

pCMV-Scn7a-3×FLAG-Neo plasmid Description

pCMV-Scn7a-3×FLAG-Neo is a plasmid suitable for protein expression in mammalian cells.

pCMV-Scn7a-3×FLAG-Neo plasmid Sequence

1 GTTAGGCGTT TTGCGCTGCT TCGCGATGTA CGGGCCAGAT ATACGCGTTG ACATTGATTA 61 TTGACTAGTT ATTAATAGTA ATCAATTACG GGGTCATTAG TTCATAGCCC ATATATGGAG 121 TTCCGCGTTA CATAACTTAC GGTAAATGGC CCGCCTGGCT GACCGCCCAA CGACCCCCGC 181 CCATTGACGT CAATAATGAC GTATGTTCCC ATAGTAACGC CAATAGGGAC TTTCCATTGA 241 CGTCAATGGG TGGAGTATTT ACGGTAAACT GCCCACTTGG CAGTACATCA AGTGTATCAT 301 ATGCCAAGTA CGCCCCCTAT TGACGTCAAT GACGGTAAAT GGCCCGCCTG GCATTATGCC 361 CAGTACATGA CCTTATGGGA CTTTCCTACT TGGCAGTACA TCTACGTATT AGTCATCGCT 421 ATTACCATGG TGATGCGGTT TTGGCAGTAC ATCAATGGGC GTGGATAGCG GTTTGACTCA 481 CGGGGATTTC CAAGTCTCCA CCCCATTGAC GTCAATGGGA GTTTGTTTTG GCACCAAAAT 541 CAACGGGACT TTCCAAAATG TCGTAACAAC TCCGCCCCAT TGACGCAAAT GGGCGGTAGG 601 CGTGTACGGT GGGAGGTCTA TATAAGCAGA GCTCTCTGGC TAACTAGAGA ACCCACTGCT 661 TACTGGCTTA TCGAAATTAA TACGACTCAC TATAGGGAGA CCCAAGCTGG CTAGCGTTTA 721 AACTTAAGCT TGGTACCGAG CTCGAATTCG CCACCatgtt gacttcccca gagccgaagg 781 gccttgtccc atttacaaca gagtcacttg aacttataga aaatcacatt gctaaaaaat 841 gcaatgaaga ccccgaagaa gaagaaggtt taaaaccaag tcgtaatcta gaagctggca 901 aaagacttcc aattccctat ggaaccctcc ctcgaggaac cgtgtcagag cccttggaag 961 atgtggatcc atactactat gttaagagaa atactttcat ggtcttaaac agaagcagag 1021 tcatcttcag gttcaatgcg gtttccatct tctgcacatt gtctcctcta aactccctca 1081 gaagagcagc tatcaaggct ttggtgcatc ccctttttcg cctgctgatt ttaatcagcg 1141 ttctcactga cagcatactt atgtgcatga gtaatctacc agaatggata ttggcaatag 1201 agaatacttt gcttgggatt tacgcatttg aaatacttgt aaaagtcatt gcaagaggta 1261 tctgggcagg ttcattttcc ttccttgggg atctttggaa ctggcttgat ttcagtgtaa 1321 ctttgttcga gctaatcaca aggttttcac ctctaagctc ctttttaatg cttaaaacta 1381 tcagaacttt ccgaattttg aagattatcc ctttgaacca cggcctgcag tctattgtga 1441 tgacactggc ccagtgtttg aagaaactat ttggtgccat tgccctagct ctgttttttc 1501 tggctgtgtt ttcactactt ggaatgggcc tcttcatggg caacctgaag cacaaatgtc 1561 tgcggtggcc agaagaaaat gaaaatgaaa cgctgcacaa cagaactgga agccttaact 1621 atagtccaga aagaataaac ttctactaca tggaaggagc gaaatatgct ctcctttgcg 1681 gcaacaggac agatgctggc cagtgtccgg aaggttatgt gtgtgtaaaa gaaggcacaa 1741 atcctgacaa tggcttcaca agttttgaca actttggctg gtcccttctt gctatgtttc 1801 gattgatgac acaggattac cctgaattac tttatcacca gatcctttat gcttcaggaa 1861 aggtctacat gatatttttt gttatgatca gtttctggtt tgccttctat ttgacaagtt 1921 tgttcttggg catactcact atgacctatg aaaaggaaaa gcagagagcc tgtgaggaat 1981 ctggaggcct tgatcccaaa tgtcaacaga cagtgaaaga actcgacgaa gaaaatgacg 2041 cagctgagat ggaaactaca caaatagaaa tgaagaaaag atccccaact tctataaaca 2101 ccacactgga tatactggaa gacactaccc tcggacacag agaagaacca gaaacatcca 2161 ggaagaaatg cccaatatgt tggcataagt ttattaaaac ctgcttcatc tggaaatgct 2221 ctccctgttg ggtaaaactg aacgagtttg ctgatagagt tataacacac ccattggctg 2281 atctttttct tgtcatctgc atcgttttaa acatatgctt cctcgcccta gaacattttc 2341 caatgagcga ggagctcagg tctctccttc acgttggaaa tttggttttt attggaattt 2401 acacaataga aatgattttg aagataatcg ctatgcatcc atatgggtat tttcaaataa 2461 gctggaatat ttttgacagt atacttgtgg ttttggagtt aacagaaatt ttactagcag 2521 atgttgaagg actagctgtt ttaataacag tcccattgat atttataaaa ctggggaagt 2581 acggaccacc atttaagagt ttgatgcgca tccttggtag ctcattgatg gccctgaaag 2641 atttggtcct gttgctctgc atattcgttt acttctctgc tgtgttcggc atgaagctgt 2701 ttggtcgaag ttacaaggat tgtgtctgcc acataaagga agactgccaa ccccaacgct 2761 ggcacatgag tgacttcctt catgcctaca tgaccgtgtt ccgaatcctc tgtggagagt 2821 ggatagagac attatgggag tgtatggagg ttgcaggcca ggcctggtgt attccttttt 2881 acatgatggt cattttaatt ggaaacttat tgatacttta cctctttgtg acattggtga 2941 gctctttcag ttactatgat gctacatcgg aagtgaacaa agaagccaaa aaccttcagc 3001 ttgccatggc aaggataaag tcgggaataa actccatgct tcttaaatta atgtgcacag 3061 aaagaagtgt tcctacagaa gcaacagacc aaatatgtga tccaagtgtt aaagagaata 3121 tttctggcca tactctttct gaactgagca acacccaaac tttcctcaga tataaggacc 3181 agagcagcag cactgagaaa actccagtga ctgaatctga gagtcaatct ctgattgcta 3241 gtcccagtgc ctctgaaact gtgccgattg cttcaggaga atctgatata gaaaatctgg 3301 ataacaagga gactcggagc aagtctggga atggaggcag taaagagaaa atgaagcagt 3361 ctagctcatc tgagtgcagc acagttgata tcgctatttc tgaagaagaa gaaatggtct 3421 atgaacatga aaagtcaaag cttcataaaa atggttatga acgcaaatct tcaactggtc 3481 aaatcagtag agaatctagg aatggaaaga tttggaaaaa catcaggaaa acttgctgca 3541 agatagtaga gaacagctgg tttgagtgtt tcattggcct ggtcactctg ctctgcacag 3601 gcactctggc tcttgaagac atatatattg atcagagaaa aaccactaaa atcttactgg 3661 aatatgcgga catgatattt gcatacatct tcattctgga gatgcttctc aagtgggtgg 3721 cttatggctt taaagccttt ttctccaaca actggtacaa actggacttt atggttgtta 3781 tcgtgttttg tcttagctta ataggcaaaa ctcgagaaga tctgaaccct ctgacatcaa 3841 taaagttcct tcgagcacta agagttctgt cgcagtttga aagaatgaag gtggtcctga 3901 gagctttgat aaaaacaacc ttacccactg tgagcgtgtt tctagtctgc ctaatgatct 3961 ggctgctttt cagtgttatt ggagtgcagt tatttgctgg caagttctat gaatgcattg 4021 acccaacaaa gggagaaaga ttccctgtat ttgaagttat gaataaaagt cagtgtgaaa 4081 aactgttatt caatgaatca atgccgtggg agaatgcaaa actgaacttt gataatgttg 4141 gaaatggttt tctttcttta ctccaagtgg caacatttaa tggatggatc agtattatga 4201 attcagcaat tgattctgtt ggtgtaaaca tgcagcccag ctttgagtac aacctctaca 4261 tgtatagtta ctttatcatc tttgttatct ttggattatt tcttcctctc tgtatgctga 4321 ttggtgttat tattcgcaat ttcaacaagc agaaaattaa gcagggagga tcaaacatct 4381 ttataacagt aaaacagaaa aaacagtacc gggccctgaa gaagctcttg tatgcagacg 4441 tccagaaacc aacaccccgc cccagaaaca aattccaagg cttccttttt gacctagtaa 4501 cacaccgtgt ctttaatgtt atcatcatac ttcttatctg tttccaagca acaaccatta 4561 tgatacaaaa ggatgagcag agtccacaaa tggaaactgc catcttctgg atgaactcca 4621 tttttgtcat gctgttcact ctggaatgca tactgaagct cactgccttc cgttgccact 4681 acttcaccag tgcatggaat gttcatgact ttatggtggt cattttctcc attacagggc 4741 tgctgctacc cttgacaata ggacaatact ttgtgcctcc ttccctggtg cagctgattc 4801 ttctctctcg agtcatccac atcctgcgtc ctgggaaagg accgaaggtg ttccatgacc 4861 tgatgcttcc cttgattctg gcgctcccag cattgctgaa cattagtctt ctcatcttcc 4921 tggtcatgtt catctacgcc atctttggaa tgtacaactt tgcctatgta aagaaagaag 4981 ccggaattaa tgatgtgtcc aactttgaga cctttggaag cagtatgctc tgtctcttcc 5041 aagttacaac gttttctggt tgggacggga tgctggatgc aattttcaac agtcagtggt 5101 ctgactgcga tcctgataaa attaatccag ggactcaggt caagggagat tgtgggagcc 5161 cttctgttgg gatttcttat tttgtcagtt acatcctcat atcatggttg atcattgtta 5221 acatgtacat tgtgttgatc atggagttct taagtattcc ttctcagaaa aaaagcagga 5281 ccttgagtga agatgacttt aggagattct tccgggtgtg gaacaggttt gaccctgata 5341 ggacccagta catagactct agcaagcttt ctgattttgc agctgctctg gatcctcctc 5401 ttttcatggc aaaaccaaac aagggccagc ttgtggccat ggatctcccc atggctgcgg 5461 gagacagaat ccactgcctc gacattttac ttgcctttac gaaaagagtg atgggaaagg 5521 atgagagggt ggagaaaatc ctttcagaga tagaatccgg gttcatgtta gcgaaccctt 5581 tcaaaatcac ttatgagccg attacaacta ctttgaaacg caaacaagag gcagtttcag 5641 caaccatcat ccagcgtgca tataaaagct accgcttaag gcaaaatgac aagaatgtat 5701 cagatactcc tgctatagat gaccgcagag atgatcttac ttctaaaggt gctcactctg 5761 gcaaaatcga ggagaaggca tctattcaga cccagattGG ATCCGACTAC AAAGACCATG 5821 ACGGTGATTA TAAAGATCAT GACATCGACT ACAAGGATGA CGATGACAAG TAGTGAGGGC 5881 CCGATATCTC GACAATCAAC CTCTGGATTA CAAAATTTGT GAAAGATTGA CTGGTATTCT 5941 TAACTATGTT GCTCCTTTTA CGCTATGTGG ATACGCTGCT TTAATGCCTT TGTATCATGC 6001 TATTGCTTCC CGTATGGCTT TCATTTTCTC CTCCTTGTAT AAATCCTGGT TGCTGTCTCT 6061 TTATGAGGAG TTGTGGCCCG TTGTCAGGCA ACGTGGCGTG GTGTGCACTG TGTTTGCTGA 6121 CGCAACCCCC ACTGGTTGGG GCATTGCCAC CACCTGTCAG CTCCTTTCCG GGACTTTCGC 6181 TTTCCCCCTC CCTATTGCCA CGGCGGAACT CATCGCCGCC TGCCTTGCCC GCTGCTGGAC 6241 AGGGGCTCGG CTGTTGGGCA CTGACAATTC CGTGGTGTTG TCGGGGAAGC TGACGTCCTT 6301 TCCATGGCTG CTCGCCTGTG TTGCCACCTG GATTCTGCGC GGGACGTCCT TCTGCTACGT 6361 CCCTTCGGCC CTCAATCCAG CGGACCTTCC TTCCCGCGGC CTGCTGCCGG CTCTGCGGCC 6421 TCTTCCGCGT CTTCGCCTTC GCCCTCAGAC GAGTCGGATC TCCCTTTGGG CCGCCTCCCC 6481 GCCTGGAAAC GGGGGAGGCT AACTGAAACA CGGAAGGAGA CAATACCGGA AGGAACCCGC 6541 GCTATGACGG CAATAAAAAG ACAGAATAAA ACGCACGGGT GTTGGGTCGT TTGTTCATAA 6601 ACGCGGGGTT CGGTCCCAGG GCTGGCACTC TGTCGATACC CCACCGAGAC CCCATTGGGG 6661 CCAATACGCC CGCGTTTCTT CCTTTTCCCC ACCCCACCCC CCAAGTTCGG GTGAAGGCCC 6721 AGGGCTCGCA GCCAACGTCG GGGCGGCAGG CCCTGCCATA GCAGATCTGC GCAGCTGGGG 6781 CTCTAGGGGG TATCCCCACG CGCCCTGTAG CGGCGCATTA AGCGCGGCGG GTGTGGTGGT 6841 TACGCGCAGC GTGACCGCTA CACTTGCCAG CGCCCTAGCG CCCGCTCCTT TCGCTTTCTT 6901 CCCTTCCTTT CTCGCCACGT TCGCCGGCTT TCCCCGTCAA GCTCTAAATC GGGGCATCCC 6961 TTTAGGGTTC CGATTTAGTG CTTTACGGCA CCTCGACCCC AAAAAACTTG ATTAGGGTGA 7021 TGGTTCACGT AGTGGGCCAT CGCCCTGATA GACGGTTTTT CGCCCTTTGA CGTTGGAGTC 7081 CACGTTCTTT AATAGTGGAC TCTTGTTCCA AACTGGAACA ACACTCAACC CTATCTCGGT 7141 CTATTCTTTT GATTTATAAG GGATTTTGGG GATTTCGGCC TATTGGTTAA AAAATGAGCT 7201 GATTTAACAA AAATTTAACG CGAATTAATT CTGTGGAATG TGTGTCAGTT AGGGTGTGGA 7261 AAGTCCCCAG GCTCCCCAGC AGGCAGAAGT ATGCAAAGCA TGCATCTCAA TTAGTCAGCA 7321 ACCAGGTGTG GAAAGTCCCC AGGCTCCCCA GCAGGCAGAA GTATGCAAAG CATGCATCTC 7381 AATTAGTCAG CAACCATAGT CCCGCCCCTA ACTCCGCCCA TCCCGCCCCT AACTCCGCCC 7441 AGTTCCGCCC ATTCTCCGCC CCATGGCTGA CTAATTTTTT TTATTTATGC AGAGGCCGAG 7501 GCCGCCTCTG CCTCTGAGCT ATTCCAGAAG TAGTGAGGAG GCTTTTTTGG AGGCCTAGGC 7561 TTTTGCAAAA AGCTCCCGGG AGCTTGTATA TCCATTTTCG GATCTGATCA AGAGACAGGA 7621 TGAGGATCGT TTCGCATGAT TGAACAAGAT GGATTGCACG CAGGTTCTCC GGCCGCTTGG 7681 GTGGAGAGGC TATTCGGCTA TGACTGGGCA CAACAGACAA TCGGCTGCTC TGATGCCGCC 7741 GTGTTCCGGC TGTCAGCGCA GGGGCGCCCG GTTCTTTTTG TCAAGACCGA CCTGTCCGGT 7801 GCCCTGAATG AACTGCAGGA CGAGGCAGCG CGGCTATCGT GGCTGGCCAC GACGGGCGTT 7861 CCTTGCGCAG CTGTGCTCGA CGTTGTCACT GAAGCGGGAA GGGACTGGCT GCTATTGGGC 7921 GAAGTGCCGG GGCAGGATCT CCTGTCATCT CACCTTGCTC CTGCCGAGAA AGTATCCATC 7981 ATGGCTGATG CAATGCGGCG GCTGCATACG CTTGATCCGG CTACCTGCCC ATTCGACCAC 8041 CAAGCGAAAC ATCGCATCGA GCGAGCACGT ACTCGGATGG AAGCCGGTCT TGTCGATCAG 8101 GATGATCTGG ACGAAGAGCA TCAGGGGCTC GCGCCAGCCG AACTGTTCGC CAGGCTCAAG 8161 GCGCGCATGC CCGACGGCGA GGATCTCGTC GTGACCCATG GCGATGCCTG CTTGCCGAAT 8221 ATCATGGTGG AAAATGGCCG CTTTTCTGGA TTCATCGACT GTGGCCGGCT GGGTGTGGCG 8281 GACCGCTATC AGGACATAGC GTTGGCTACC CGTGATATTG CTGAAGAGCT TGGCGGCGAA 8341 TGGGCTGACC GCTTCCTCGT GCTTTACGGT ATCGCCGCTC CCGATTCGCA GCGCATCGCC 8401 TTCTATCGCC TTCTTGACGA GTTCTTCTGA GCGGGACTCT GGGGTTCGCG AAATGACCGA 8461 CCAAGCGACG CCCAACCTGC CATCACGAGA TTTCGATTCC ACCGCCGCCT TCTATGAAAG 8521 GTTGGGCTTC GGAATCGTTT TCCGGGACGC CGGCTGGATG ATCCTCCAGC GCGGGGATCT 8581 CATGCTGGAG TTCTTCGCCC ACCCCAACTT GTTTATTGCA GCTTATAATG GTTACAAATA 8641 AAGCAATAGC ATCACAAATT TCACAAATAA AGCATTTTTT TCACTGCATT CTAGTTGTGG 8701 TTTGTCCAAA CTCATCAATG TATCTTATCA TGTCTGTATA CCGTCGACCT CTAGCTAGAG 8761 CTTGGCGTAA TCATGGTCAT AGCTGTTTCC TGTGTGAAAT TGTTATCCGC TCACAATTCC 8821 ACACAACATA CGAGCCGGAA GCATAAAGTG TAAAGCCTGG GGTGCCTAAT GAGTGAGCTA 8881 ACTCACATTA ATTGCGTTGC GCTCACTGCC CGCTTTCCAG TCGGGAAACC TGTCGTGCCA 8941 GCTGCATTAA TGAATCGGCC AACGCGCGGG GAGAGGCGGT TTGCGTATTG GGCGCTCTTC 9001 CGCTTCCTCG CTCACTGACT CGCTGCGCTC GGTCGTTCGG CTGCGGCGAG CGGTATCAGC 9061 TCACTCAAAG GCGGTAATAC GGTTATCCAC AGAATCAGGG GATAACGCAG GAAAGAACAT 9121 GTGAGCAAAA GGCCAGCAAA AGGCCAGGAA CCGTAAAAAG GCCGCGTTGC TGGCGTTTTT 9181 CCATAGGCTC CGCCCCCCTG ACGAGCATCA CAAAAATCGA CGCTCAAGTC AGAGGTGGCG 9241 AAACCCGACA GGACTATAAA GATACCAGGC GTTTCCCCCT GGAAGCTCCC TCGTGCGCTC 9301 TCCTGTTCCG ACCCTGCCGC TTACCGGATA CCTGTCCGCC TTTCTCCCTT CGGGAAGCGT 9361 GGCGCTTTCT CAATGCTCAC GCTGTAGGTA TCTCAGTTCG GTGTAGGTCG TTCGCTCCAA 9421 GCTGGGCTGT GTGCACGAAC CCCCCGTTCA GCCCGACCGC TGCGCCTTAT CCGGTAACTA 9481 TCGTCTTGAG TCCAACCCGG TAAGACACGA CTTATCGCCA CTGGCAGCAG CCACTGGTAA 9541 CAGGATTAGC AGAGCGAGGT ATGTAGGCGG TGCTACAGAG TTCTTGAAGT GGTGGCCTAA 9601 CTACGGCTAC ACTAGAAGGA CAGTATTTGG TATCTGCGCT CTGCTGAAGC CAGTTACCTT 9661 CGGAAAAAGA GTTGGTAGCT CTTGATCCGG CAAACAAACC ACCGCTGGTA GCGGTGGTTT 9721 TTTTGTTTGC AAGCAGCAGA TTACGCGCAG AAAAAAAGGA TCTCAAGAAG ATCCTTTGAT 9781 CTTTTCTACG GGGTCTGACG CTCAGTGGAA CGAAAACTCA CGTTAAGGGA TTTTGGTCAT 9841 GAGATTATCA AAAAGGATCT TCACCTAGAT CCTTTTAAAT TAAAAATGAA GTTTTAAATC 9901 AATCTAAAGT ATATATGAGT AAACTTGGTC TGACAGTTAC CAATGCTTAA TCAGTGAGGC 9961 ACCTATCTCA GCGATCTGTC TATTTCGTTC ATCCATAGTT GCCTGACTCC CCGTCGTGTA 10021 GATAACTACG ATACGGGAGG GCTTACCATC TGGCCCCAGT GCTGCAATGA TACCGCGAGA 10081 CCCACGCTCA CCGGCTCCAG ATTTATCAGC AATAAACCAG CCAGCCGGAA GGGCCGAGCG 10141 CAGAAGTGGT CCTGCAACTT TATCCGCCTC CATCCAGTCT ATTAATTGTT GCCGGGAAGC 10201 TAGAGTAAGT AGTTCGCCAG TTAATAGTTT GCGCAACGTT GTTGCCATTG CTACAGGCAT 10261 CGTGGTGTCA CGCTCGTCGT TTGGTATGGC TTCATTCAGC TCCGGTTCCC AACGATCAAG 10321 GCGAGTTACA TGATCCCCCA TGTTGTGCAA AAAAGCGGTT AGCTCCTTCG GTCCTCCGAT 10381 CGTTGTCAGA AGTAAGTTGG CCGCAGTGTT ATCACTCATG GTTATGGCAG CACTGCATAA 10441 TTCTCTTACT GTCATGCCAT CCGTAAGATG CTTTTCTGTG ACTGGTGAGT ACTCAACCAA 10501 GTCATTCTGA GAATAGTGTA TGCGGCGACC GAGTTGCTCT TGCCCGGCGT CAATACGGGA 10561 TAATACCGCG CCACATAGCA GAACTTTAAA AGTGCTCATC ATTGGAAAAC GTTCTTCGGG 10621 GCGAAAACTC TCAAGGATCT TACCGCTGTT GAGATCCAGT TCGATGTAAC CCACTCGTGC 10681 ACCCAACTGA TCTTCAGCAT CTTTTACTTT CACCAGCGTT TCTGGGTGAG CAAAAACAGG 10741 AAGGCAAAAT GCCGCAAAAA AGGGAATAAG GGCGACACGG AAATGTTGAA TACTCATACT 10801 CTTCCTTTTT CAATATTATT GAAGCATTTA TCAGGGTTAT TGTCTCATGA GCGGATACAT 10861 ATTTGAATGT ATTTAGAAAA ATAAACAAAT AGGGGTTCCG CGCACATTTC CCCGAAAAGT 10921 GCCACCTGAC GTCGACGGAT CGGGAGATCT CCCGATCCCC TATGGTCGAC TCTCAGTACA 10981 ATCTGCTCTG ATGCCGCATA GTTAAGCCAG TATCTGCTCC CTGCTTGTGT GTTGGAGGTC 11041 GCTGAGTAGT GCGCGAGCAA AATTTAAGCT ACAACAAGGC AAGGCTTGAC CGACAATTGC 11101 ATGAAGAATC TGCTTAGG

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Caution:
1.This product is FOR RESEARCH USE ONLY!
2.The item is lyophilized form, Please take the powder plasmid by centrifugation at 5000rpm/min for 1min. Add 20μl ddH2O in to the tube of plasmid.Take 2ul plasmid into 100 μ L corresponding competent cell, centrifugal then full coating on plate.
3.Shipping temperature is 2-8???