competent cell,cloning vector, transformation

[anxenz88]

Did someone know how the CaCl2 can make the membrane cell become permeable to DNA in transformation?

[osama al refa'e]

maybe this link will hilp

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[aftabac]

Hi
its nice link with lot of more informations on different topics.....

regards
aftab

[Barbienidoo]

Hi,

for transformation we make competent cells to enable them for uptake of foreign DNA.For this purpose we use different methods of which one is chemical method i.e using CaCl2 which gives shocks to membrane of the cells rendering them fragile thus helping in the uptake of foreign genome.

[Liquid Nitrogen]

Hi Obama, the link posted by you is not opening, can you check it again?

[osama al refa'e]

hi
Obama , hh, nice one , dear liquid nitrogen
i checked the link more than one time and it worked with me
let me know if it work with you ok?>>> have a nice day

[magdziec]

Hi, maybe someone of you could help me to understand the reason of my problem. I want to clone 2 genes of heme proteins, so I added the second gene to the first one, present on the plasmid. I didn't remove the stop codon of the first gene because i put the ribosome binding site before the second one. In this way I would like to have 2 separated proteins. As heme proteins are transfered to periplasm I don't need any tag. I checked also the presence of right sequences in the plasmid amplificated by PCR. SDS-PAGE of bacteries growth showed the right size of the proteins which should be red but they are white. Did somebody do something like this?