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COMPETENT CELL PREPARATION


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#1 marine

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Posted 28 August 2009 - 12:52 AM

I am using CaCl2 for preparation of competent cells in my lab but I have read other protocols also in which MgCl2 is used in place of CaCl2 B) . Can anyone tell me that which is the better salt for competent cell preparation and also what is the diffrence in their functioning? Also does anyone know the exact reason for adding these salts , all I know is that they bind to the DNA backbone?

#2 pito

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Posted 28 August 2009 - 04:34 AM

When I prepare competent cells I use both: MgCl2 and CaCl2.

I think it all depends on what protocol you use.

What protocol do you use?

And why do you need to use the MgCl2 and CaCl2 , I do not remember the exaxt reason, but I can check it this weekend in one of my textbooks.

If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.


#3 marine

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Posted 31 August 2009 - 12:40 AM

When I prepare competent cells I use both: MgCl2 and CaCl2.

I think it all depends on what protocol you use.

What protocol do you use?

And why do you need to use the MgCl2 and CaCl2 , I do not remember the exaxt reason, but I can check it this weekend in one of my textbooks.



#4 marine

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Posted 31 August 2009 - 12:44 AM

When I prepare competent cells I use both: MgCl2 and CaCl2.

I think it all depends on what protocol you use.

What protocol do you use?

And why do you need to use the MgCl2 and CaCl2 , I do not remember the exaxt reason, but I can check it this weekend in one of my textbooks.


I use the CaCl2 protocol, but I want to see if using MgCl2 or both CaCl2 n MgCl2 gives better results.

#5 pito

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Posted 31 August 2009 - 02:53 AM

When I prepare competent cells I use both: MgCl2 and CaCl2.

I think it all depends on what protocol you use.

What protocol do you use?

And why do you need to use the MgCl2 and CaCl2 , I do not remember the exaxt reason, but I can check it this weekend in one of my textbooks.


I use the CaCl2 protocol, but I want to see if using MgCl2 or both CaCl2 n MgCl2 gives better results.


Do you have protocols that involve the use of both MgCl2 and CaCl2 ? If not, I can give you the protocol I use.

And how do you know your protocol is not good?

If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.


#6 lizzy

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Posted 03 September 2009 - 07:18 PM

I used Cacl2 to prepare competent cells. I think the Cacl2 protocal can give us a good result and there is no need to prepare competent cells using Mgcl2.

#7 Chimp

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Posted 25 September 2009 - 02:39 AM

My experience with CaCl2 alone is very good. Never tried anything else. The important thing in my experience is using FRESH cells. If you prepare them the same day, which is pretty simple, they (BL21) transform brilliantly!




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