One Shot TOP10 Cells
Posted 18 April 2011 - 12:25 AM
I have discovered a box of One Shot TOP10 competent cells deep within our deep freezer. So I am very curious about how I can use this.
I have looked up a manual for these cells and it says that "TOP 10 Cells do not require IPTG to induce expression from the Lac promoter".
So my question is- if I have a plasmid from the pET system under T7 promoter control, I still need to add IPTG to induce my target protein right??
I am also looking at the genotype for this. What does StrR mean? streptomycin resistance?
Posted 18 April 2011 - 02:14 AM
Hola yes streptomycin resistance, but this is a subcloning strain that only need Xgal for blue/white selection, in fact after transformation it says check the insert presence by PCR or whatever.This is an efficient strain to transform with ligation mixtures equivalent to JM103or DH5 alpha as pDNA said to you in your question before. Buena suerte LB
http://www.chem.eng.osaka-u.ac.jp/~inoue-tken/ja/staffs/matsumura/OneshotTOP10_man.pdf was the link I got the info from
Edited by protolder, 18 April 2011 - 02:40 AM.
Posted 18 April 2011 - 02:39 PM
Thank you for your reply. Has anyone had experience with using this particular strain for expression?
Would it be useful for that purpose, or it it more just a cloning strain for making plasmids?
Posted 18 April 2011 - 06:43 PM
Posted 18 April 2011 - 08:56 PM
Posted 18 April 2011 - 10:42 PM
Thanks for this.