Transformation-Min plasmid reqd - (Mar/26/2007 )

Hello All,

I will receive one plasmid from other lab and I just got an email gtom them that they are sending only 5µl and he concentration is about 100ng/µl. Don't ask me why they sent only 5µl but now I will have to transform it into DH5a bacteria.

I would like to know how much minimum plasmid amount I should use for this transformation. I usually use 50 or 100µl aliquot of competent DH5a cells.

Many thanks for your suggestions.

(P.S. - I don't want to use all as if something goes wrong, I will be in trouble and we will receive this plasmid from the other lab after a lot of hassle)

Hello Jiang M!

Usually in our lab we transform 50-100 ng of plasmid into a 50µl aliquot of DH5-alpha.

And after all you only need one conlony to prepare a new culture... So don't worry - I guess 1µl might be enough.

Greetings,
Chakchel

You could use 100 pico grams to transform and get a lot of colonies. I use something around 1ng to retransform.

What they are sending is more than sufficient.

Thank you both of you for quick reply. I am very relieved.