cloning in yeast two hybrid system vector

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Gokila Devi Mat...
Gokila Devi Mathialagan's picture
cloning in yeast two hybrid system vector

I have been trying to clone in the yeast two hybrid system vectors(pGBKT7 & pGADT7). My insert is of 52 bp (which we designed as forward and reverse primer with ECOR1 site -suspended in autoclaved double distilled water & annealed in 100 degree Celsius boiling water).I ligated the vector -10 nanogram and insert -300 picomole,overnight,16 degree celsius.but , i couldnot get the clone.help me to troubleshoot.