it worked! hahahahaha - (Sep/24/2008 )
sorry I just had to share this, after a long hard week full of optimizations and mistakes, my western blot worked and its looks adorable if i may say so myself....................
so great...............THANK YOU FOR ALL THE TIPS AND SUPPORT!!!!!!! hurray to this forum!!!!!!!
Congratulations!!! Well done! I am happy for you knowing how hard it can be to establish a new antibody / WB, great! Enjoy your triumph
Great !!! Its always nice when things work.
Have a great Week !!!
uhm......so my cloning didnt work today booooooooooooooooooo haha but since I had one positive result this week I am not giving up, my plates looked like fields of bacteria were growing on them, so I guess this could mean religated vector OR maybe my ampicillin isnt working. Religated vector is unlikely, I mean I used a 0.7% gel to run about 600 ng of vector and I let it run for aaaaaaaages before cutting it out. So i asked this girl in the other subgroup if I could have one of her plates and I am replating what I have left of my transformation and seeing what happens.
I will order a CIP today though, a CIP and a kinase for my oligos, and I will digest fresh vector.
Just in case I mess up is there anything one really SHOULDNT do when phosphorylating oligos and dephosphorylating vector? Anyone? lol