I am using XL-10 gold cells for transformation following PCR. Normally I use the B-mercaptoethanol provided with the cells by Agilent. However, I have run out of it, and I was wondering if anybody has any experience using reagent grade B-ME with the XL-10 gold cells? Is there a difference?
Molecular biology grade BME works fine in my hands with chemically competent XL-10.
Daniel: how much of the molecular biology grade 2-mercaptoethanol do you use? I mean, if you take it straight out of the bottle of concentrated (~14M) stock, how much is needed; alternatively, how much final concentration do you use?
I would recommend a 1:10 dilution of BME stock into water. Protocols do vary a bit, probably anything between a 1:10 and a 1:25 dilution should work.
BME is not compulsory for the transformation, however, it can increase the efficiency of the transformation by inactivating surface nucleases. So, even if you perform transformation without using BME, it should at least work.
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