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hi ,
my supervisor just asked me to add 0.03 % tween80 to my bacterial culture which i induced by IPTG , he believes that it helps the proteins trapped in membrane be secreted easily so i will be able to extract them from the supernatant after centrifuging bacterial cells .
has any body heard any thing about this ? what is tween for ? doesn't it have negative effects on my cells and my recombinant protein ?!
thanks in advance ...
Mina
Topics I've Started
plasmid extraction troubleshooting,please help !!
02 June 2013 - 09:07 AM
hi every one
i need help with plasmid extraction . my plasmid is PET21a and as u know it's an expression vector so it gives low concentrations in extraction ( or maybe i am the problem !!
) . any body has any suggestions to get higher yields ??
i truly appreciate ur help
i need help with plasmid extraction . my plasmid is PET21a and as u know it's an expression vector so it gives low concentrations in extraction ( or maybe i am the problem !!
) . any body has any suggestions to get higher yields ??i truly appreciate ur help
doing PCR using PCR product , help plz !!
04 February 2013 - 06:03 AM
hi every one
let me get straight to the point . i am running out of my template DNA so i decided to use the pcr product of it as a template for another pcr .
so i used my main template DNA for the pcr . after my pcr was done i used 5 microliter of it and added it to a new pcr kit , then i added 15 microliter distilled water , and 1 micoliter of each primer . and ran the same program that had given a very good pcr product in the first place .... but , it didnt work !!!
and i a m really worried plz guys if u know any thing that could help me , do tell !
regards
Mina
let me get straight to the point . i am running out of my template DNA so i decided to use the pcr product of it as a template for another pcr .
so i used my main template DNA for the pcr . after my pcr was done i used 5 microliter of it and added it to a new pcr kit , then i added 15 microliter distilled water , and 1 micoliter of each primer . and ran the same program that had given a very good pcr product in the first place .... but , it didnt work !!!
and i a m really worried plz guys if u know any thing that could help me , do tell !regards
Mina
