Once V is added into Al alloys containing Ti and B, it it likely that V will react with B to form stable VB2 in the alloy. This may lead to change in the fluidity of alloy.
check this web: www.diva-portal.org/smash/get/.../FULLTEXT01.pdf
However, did a transformation a month before with a pGEX6P2 vector into DH5 alpha and got colonies, so I feel transformation is working. Taking that forward...that transformation is working. it's something to do with ligation.
repeated ligation: this time overnight ligation: 4C, 18hours, 1:3 (V:I), 20uL reaction, 5% ligase -> Did two transformations: T1: 1uL into 50uL cells -> no colonies T2: 3uL into 50uL cells -> no colonies