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Problem to do the library. Template Switch.
Hi everyone.
I have a problem with my template switch (TS) to make the library. The TS doesn't stick to the transcript of cDNA.
I'm using an enzyme Superscript II with TDT activity, and theoretically, it would work. I using the standard protocol of Superscript II, and another second option to make the cDNA synthesis in two steps, first only cDNA synthesis and after second to complete TS stick. Neither of these two options works and I repeat these experiments many times.
Any suggestions to find the problem? If you need more details to think about the problem only ask me.
Thanks for your attention.
I have a problem with my template switch (TS) to make the library. The TS doesn't stick to the transcript of cDNA.
I'm using an enzyme Superscript II with TDT activity, and theoretically, it would work. I using the standard protocol of Superscript II, and another second option to make the cDNA synthesis in two steps, first only cDNA synthesis and after second to complete TS stick. Neither of these two options works and I repeat these experiments many times.
Any suggestions to find the problem? If you need more details to think about the problem only ask me.
Thanks for your attention.