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These three DNA fragments with overlapping sequences were individually inserted into the pMD18-T vector (Takara, http://www.takara.com.cn/) to produce the 7179 bp fragment.
The XbaI XhoI fragment of FeaB, AldB or AldH from pDG3, pDG4 or pDG5 was individually inserted into NheI and XhoI sites of pDG30 to give pDG6, pDG7 and pDG8.
Equal weights of each film were individually inserted in test tubes, each containing 10 ml of sterile Mueller Hinton (MH) broth (composed of g/l: beef extract, 2.0; casein hydrolysate, 17.5 and starch 1.5; pH 7.3 ± 0.2).
Amplified sequences were then individually inserted, in both orientations, at the BamHI unique site of pIRT2Δ.
The respective target sequences were individually inserted into the 3′ UTR of a plasmid-located Renilla luciferase gene.
The coding sequences were individually inserted into pHEX2 vector under the control of 35S promoter using the LR Clonase II Kit (Invitrogen).
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The tiles were readout individually via inserted wavelength-shifting fibres.
Animals were tested individually by inserting one by one into the experimental tank.
These peptide sequences were successfully inserted individually as well as all together in a multivalent strategy.
In the first series, one, three, five and seven blastomeres were biopsied from 2-, 4-, 6- and 8-cell embryos respectively, and inserted individually into empty ZP recipients, leaving the remaining blastomere within its original ZP.
Each oligonucleotide tag was inserted individually into the blunted KpnI site of p TnMod-OΩ (Fig. 1).
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