In my Lab we are constructing an RNAi vector ( pPOTLX-1 ) and every time we test LR recombination results using PCR primers and NPT II primers to test the presence of our inserted gene we get the beautiful bands ( Fig 1).Thereafter we use restrictions sites( XbaI and XhoI) of our pHELLSGATE vector since its the one we are using as our Carrier vector to check the presence of inserted sequences ( Fig 2) this test also show some bands that confirms the presences of the inserts but when we go further to confirms the presence of our inserts by sequencing the sequencing company tells us that our LR recombination it just a failure so is there any other methods to identify and verify our RNAi vector constructs or we should just go on with our experiment?
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