SURE E. coli strain - How make competents cells? (Jun/26/2007 )
Hello!
I would like to know if someone can send me - or even if someone knows - a specific protocol to competent SURE E. coli strain. I get problems to do it!
Now, I'll try the "Inoue method for Ultra-competent cells". But I dont know if work well to this strain.
I'm trying to cloning small oligos to RNAi.
If someone can help me, I'll be very thankfull!
Cleidsonn
-Cleidsonn-
QUOTE (Cleidsonn @ Jun 27 2007, 11:21 AM)
Hello!
I would like to know if someone can send me - or even if someone knows - a specific protocol to competent SURE E. coli strain. I get problems to do it!
Now, I'll try the "Inoue method for Ultra-competent cells". But I dont know if work well to this strain.
I'm trying to cloning small oligos to RNAi.
If someone can help me, I'll be very thankfull!
Cleidsonn
I would like to know if someone can send me - or even if someone knows - a specific protocol to competent SURE E. coli strain. I get problems to do it!
Now, I'll try the "Inoue method for Ultra-competent cells". But I dont know if work well to this strain.
I'm trying to cloning small oligos to RNAi.
If someone can help me, I'll be very thankfull!
Cleidsonn
I would suggest that you make electrocompetent cells since it is easier and need no special trick. Make sure that you did use the correct antibiotic when you plate and grow the bacteria though so that you get the correct strain.
-Almasy-
QUOTE (Cleidsonn @ Jun 27 2007, 11:21 AM)
Hello!
I would like to know if someone can send me - or even if someone knows - a specific protocol to competent SURE E. coli strain. I get problems to do it!
Now, I'll try the "Inoue method for Ultra-competent cells". But I dont know if work well to this strain.
I'm trying to cloning small oligos to RNAi.
If someone can help me, I'll be very thankfull!
Cleidsonn
I would like to know if someone can send me - or even if someone knows - a specific protocol to competent SURE E. coli strain. I get problems to do it!
Now, I'll try the "Inoue method for Ultra-competent cells". But I dont know if work well to this strain.
I'm trying to cloning small oligos to RNAi.
If someone can help me, I'll be very thankfull!
Cleidsonn
the protocol to make the SURE cells is the same as any other competent cells. I agree It can be a bit more challenging than normal cells.
Good Luck !!!
-scolix-
What is SURE cells so special about it?
Isnt it to make competent cells, it really depends on whether it is electrocompetent or chemical competent, right?
-timjim-
QUOTE (timjim @ Jun 29 2007, 07:27 AM)
What is SURE cells so special about it?
Isnt it to make competent cells, it really depends on whether it is electrocompetent or chemical competent, right?
Isnt it to make competent cells, it really depends on whether it is electrocompetent or chemical competent, right?
SURE cells prevent recombination and if your are cloning any shRNA sequences or viral vectors with repeat sequences, using these cells prevents any sort of recombination and helps stabilize the vector and not lose parts of the vector.
-scolix-