2 replies to this topic

[Anusha]

Hi. I have 20ul of precipitated vector that we obtained from another lab. i wish to amplify and have more of the vector. Can  just transform it with competent Ecoli cells and purify doing a miniprep and then a Maxi? or is there any other quick technique to obtain more of the vector?

please help.

thanks

[Dr Teeth]

Dilute some of the vector to 1-2 ng/ul and transform in competent cells and perform a maxiprep.  When setting up your culture for the maxiprep, follow the appropriate protocol based on the plamid copy number.  Confirm the identity of your plasmid by restriction digestions and sequencing!  Many labs have sent me plasmids that were not what they were supposed to be or had mutations.  To save yourself time, energy, and resources, SEQUENCE any plasmid sent to you by anyone before you test it experiementally!

Edited by Dr Teeth, 23 September 2009 - 10:52 AM.

[Anusha]

Thanks Dr. teeth on that suggestion

Dr Teeth, on Sep 23 2009, 12:50 PM, said:

Dilute some of the vector to 1-2 ng/ul and transform in competent cells and perform a maxiprep.  When setting up your culture for the maxiprep, follow the appropriate protocol based on the plamid copy number.  Confirm the identity of your plasmid by restriction digestions and sequencing!  Many labs have sent me plasmids that were not what they were supposed to be or had mutations.  To save yourself time, energy, and resources, SEQUENCE any plasmid sent to you by anyone before you test it experiementally!

Thanks