SuperScript® Mouse 13.5 Day Embryos cDNA Library (Male, C57BI/6J) - FAQs

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2 个常见问题解答

What is the best way to prepare a plasmid prep of an Invitrogen premade library in order to get the highest representation of clones for transfection of mammalian cells?

The key to minimizing a change in library representation is to put the library through as few cell divisions as possible. Initiate a culture with as much of the frozen library as possible. Growing up the library from a very small number of starting cells is not ideal since a large number of cell divisions will be required to reach the appropriate density. Do not amplify the library because this will also change the representation.

Which promoter, T7 or SP6, should be used for an in vitro transcription/translation of clones in the pCMVSport vectors that were used to generate Invitrogen premade cDNA libraries?

The cDNA insert is cloned in the Not I/Sal I sites of the pCMVSport vectors. The Sal I site is the 5' end of the sense strand. Therefore, in order to make an in vitro transcribed/translated product from the sense strand, one would have to use the SP6 promoter.