The PCR for the amplification was composed of 10 µL Q-solution, 5 µL 10 × KOD buffer, 5 µL dNTPs (2 mM each), 2 µL MgSO4 (25 mM), 1.3 µL forward primer CYP2C19_HindIII_fwd (5′-AAGAGGAGaagcttACCATGGATCCTTTTGTGGTCCTTG-3′), 1.3 µL reverse primer ...
Th,D Panchal,L Tawk,S Thiberge,TG Carvalho,JC Barale 展开 摘要: We describe here a highly efficient procedure for conditional mutagenesis in Plasmodium. The procedure uses the site-specific recombination FLP-FRT system of yeast and targets the pre-erythrocytic stages of the rodent Plasmodium ...
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Dev79,STVY RVRE,We Rob Rave,Calculon,Austin Speed,Shamanga,MC Elixir,K-Rai,Spotovsky,Frenquency,Count Vanderhoff,Alexandre,Cristina,Illexxandra,DJ Ends,JTRA,Jah Bliddie,Subshell,Hi$to,PUFF.Magic,Dom Corleone,Homesick,DBK Teklife,DJ Flp ...
R2: CTTTCATCAATTGTGGAAGA. 2.3 Regulatory efficiency calculations Ep30TLK was cultured on LB medium supplemented with KAN + AMP + IPTG at 37°C. Samples of 50 cultures of 100 µL each were mounted on glass slides. Ep30TL competent cells were negative control without p30TK. Prepared sli...
TG ×WT 这里说的转基因,并非广义上的所有的基因修饰,而是特指外源基因随机地整合到小鼠染色体上获得的随机插入转基因小鼠。 通过受精卵显微注射的方式,通常我们会获得很多个带有外源基因插入的首建鼠(Founder)。每一个首建鼠,外源基因插入的位置或次数都是不同的。所以每个首建鼠都应该自 ...
经过限制酶酶切及部分测序鉴定其结构正确后 ,将线性化了的打靶载体以电穿孔法导入ES细胞内 ,经G418和 6 -TG双药筛选和分子鉴定 ,得到了 2个在HPRT位点整合有FLP重组酶"交换盒"F Neo F3结构的双交换重组ES细胞克隆 ,为建立基于FLP重组酶介导的盒式交换的高效、定点转基因体系创造了条件doi:CNKI:SUN:SHWU....
[0039]aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgaggcc ttcgctaatg 660 [0040]cgggaaagct cttattcggg tgagatgggc tggggcacca tctggggacc ctgacgtgaa 720 [0041]gtttgtcact gactggagaa ctcgggtttg tcgtctggtt gcgggggcgg cagttatgcg 780 [0042]gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg cc...
(H2) as well as the 20 nt sequence 5'-TGTAGGCTGGAGCTGCTTCG-3' (P2) which is complementary to sequence 3' of the spectinomycin resistance gene-rpsLNgcassette. A PCR amplicon was generated using these primers together with pRSM2832 as the template. Amplification was performed in a 50 μ...
TGG GGC TG.3 。利用 Fl /R1 对转化 子进行 PCR 鉴定。反应体积为 2O l :50 pmol / l 引物各 1 l, 10 ×反应 缓冲液 2 l ,dNTPs 1 ,MgC12 2 tzl ,Taq DNA 聚合酶 1 l。反应条件 :94℃变性 5 min,94℃ 60 S,56℃ 45 s,72oC 90 s,共 30 个循环 。扩增产物经 ...