Can I use TRIzol® or other phenol-chloroform based extractions for RNA isolation?
We do not recommend the use of TRIzol® or similar methods as any carry over of organic solvents may inhibit downstream enzyme activity. If using TRIzol extracted RNA, we recommend using a column-based purification of the RNA prior to input into the kit.
Can I use carrier RNA during RNA isolation?
We do not recommend the use of carriers during RNA isolation. If a carrier is required, please contact Technical Support for more information.
Do I need to use high quality total RNA?
Yes. Use of lower quality RNA may result in poor performance. One approach to determining RNA quality is the Agilent Bioanalyzer’s RNA Integrity Number (RIN). Clean RNA with a RIN score greater than 7 should amplify well.
How much total RNA do I need for amplification?
We recommend staying within the range of 500 pg to 50 ng total RNA starting material. Amounts greater than 50 ng may produce variable results.
Can the Crescendo cDNA Synthesis for qPCR kit amplify DNA?
The Crescendo cDNA Synthesis for qPCR kit is designed to amplify RNA, not DNA.
Can I use the Crescendo cDNA Synthesis for qPCR kit on bacterial RNA samples?
The Crescendo cDNA Synthesis for qPCR kit theoretically will work with some bacterial RNAs. However, the kit has not been optimized for this purpose.
Are there any tissues that will not work with the Crescendo cDNA Synthesis for qPCR kit?
We have not encountered any good quality, clean RNA samples that will not work with the Crescendo cDNA Synthesis for qPCR kit.