How much i add amipicillin and kanamycin into pQE30 (Qiagen) contains cloned fragment and transformed into M15 ( (pREP4) cells to do miniprep?

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Dominique Liger

Hi there,

M15(pREP4) is an expression strain. You may use 25µg/mL kanamycin and 100µg/mL ampicillin for selection. For pQE30 plasmid propagation for miniprep, use your usual cloning strain (XL1, DH5...) with 100µg/mL ampicillin.

For info:

http://www.chem.uky.edu/courses/che554/6_AffinityChro/QIAexpressionist.pdf

Syeda Mahreen Hassan

it means i donot need to add kanamycin just i have to add amipllicin

As pQE30 selection marker is AmpR then you need ampicillin for selection.