SuperScript® Plus Indirect cDNA Labeling System with Alexa Fluor® Dyes - FAQs

查看更多产品信息 SuperScript® Plus Indirect cDNA Labeling System with Alexa Fluor® Dyes - FAQs (L101406)

2 个常见问题解答

Can the amino-labeled DNA generated with your SuperScript Indirect Labeling kit be ethanol precipitated and stored overnight? Do you know if the amino-labeled DNA will be stable over that time?

Yes, the amino-labeled cDNA should be stable if stored precipitated in ethanol at -20 degrees C.

Can I use the SuperScript Indirect cDNA Labeling System for bacterial samples?

We have not used bacterial RNA for the validation of the SuperScript Indirect cDNA Labeling System, however the kit should work reasonably well with few modifications to the protocol.

For total RNA samples, Oligo(dT)20 primers should be used. As the manual suggests, random hexamers should be used only with mRNA or with the RNA ladder, which is included as a control. However, in the case of bacterial RNA, longer random primers should be used instead of the random hexamers included in the kit. 18mers should work well, although the temperature of reverse transcription may need to be slightly adjusted in order to obtain optimal cDNA synthesis. The reason for using longer random primers is that the rRNA in bacterial samples will also be used as template for reverse transcription, and the target cDNA will be more complex than the corresponding cDNA primed with the Oligo(dT)20. Therefore, when doing hybridizations, you can expect some degree of cross-hybridization to occur. Nevertheless, the use of long random 18mers is common practice among microarray users.