A modified T-vector for simplified assembly of hairpin RNAi constructs
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- Luo, K., Harding, S.A. & Tsai, CJ. Biotechnol Lett (2008) 30: 1271. doi:10.1007/s10529-008-9673-x
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We describe a modified T-vector, pGFPm-T, for direct cloning of RT-PCR products to generate bidirectional restriction fragments for assembly of hairpin-containing RNAi vectors in the popular pFGC and pGSA binary vector backbone. Green fluorescence protein (GFP) is used as a visual reporter for direct selection of recombinants under UV illumination. The simplified cloning process enables a seamless workflow from candidate gene selection and RT-PCR verification to inverted repeat cloning, using a single pair of gene-specific primers.