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| Dear All, I am planning to clone a gene of interest in qiagen expression system, using their vector pQE30 UA, in which directly the PCR amplified product can be cloned, just like the good old TA cloning. The design of primer should be in such a way that the start codon should form the first 3 nucleotide of the insert DNA. There can be no manipulations there! I have thus thought of taking 1st 23 nucleotides The problem is with the GC content of this primer. GC is just about 35%. I can manipulate the reverse primer since it is N terminal His tagging vector, therefore the ATG has to be in frame with the vector's other cis elements. Any warnings that I should be considering in particular? -- -- Yoginee Budhkar Senior Research Fellow Food Engineering and Technology Department Institute of Chemical Technology Matunga, Mumbai 400019 Contact: +91 9223355677 |
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| designing , expression , primer , vector |
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