Bacteria harvesting and lysis - (Aug/31/2006 )

Dear all,

I want to express GST-fusion protein in JM109, but, i get a problem here that the protocol of cell lysis i used can not lyse the cell very well. Dose anyone can share your protocol working well for cell lysis without sonication?

Thanks for help!!

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we use a french cell press to lyse cells in a lysis buffer containing hepes, imidazole and protease inhibitors. this works well for JM109s and M15s. cell presses are messy and not easy to use, but they get the job done. if you don't have access to a cell press you could try sonicating smaller volumes of cell suspension. make sure your lysis buffer is made fresh! hope this helps.

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I use a modified lysis buffer for my pc12 cell line with protease inhibitor. I let my cell sit on ice for 10min in the lysis buffer than I centrifuge. It's been working well. Let me know if you want the protocol.

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Dear jmo,

Thanks for your reply. Can you send the protocol to my email box "sue_mei2002@yahoo.com.tw"?
Thank you again!

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try bugbuster from novagen, add it to cells incubate for 10mins and its done


you will never need anything else!!!!!!



enjoy

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