Produce amplified amounts of cDNA from precious, limiting cell samples without bias
- Perform RNA amplification directly from cultured cell lysates followed by cDNA synthesis
- Amplify with this high-fidelity, linear RNA amplification process that preserves the relative transcript abundance of the sample.
- Get more data out of precious samples – use less RNA for more RT-qPCR reactions
- Readily and reproducibly detect even low-abundance transcripts from as little as one cell.
- No need to purify RNA: Perform RNA amplification and cDNA synthesis reactions directly from cell lysates.
- Collect samples less often and archive cDNA for future use, saving time and effort.
Applications
- Amplification of mRNA directly from a cell lysate, without the need for purifying RNA, and conversion of the amplified RNA to cDNA.
- Significantly increasing the number and sensitivity of qRT-PCRs from very small numbers of cells.
- Generation of large amounts of cDNA from the mRNA of very small samples for archival purposes.
Figure 1 (click to enlarge). An overview of the procedure for the MessageBOOSTER™ cDNA Synthesis from Cell Lysates Kit. A kit reaction amplifies the poly(A) RNA (mRNA) directly from a crude cell lysate without the need for RNA purification. The amplified RNA is then reverse-transcribed to cDNA that can be diluted up to 1,000-fold for qPCR.
Figure 2. A MessageBOOSTER™ Kit reaction produces sufficient cDNA from a single-cell lysate for thousands of sensitive qPCRs. qPCR was performed using undiluted (red), 1:10 diluted (green), 1:100 diluted (blue), and 1:1,000 diluted (purple) cDNA from a lysate of a single NRK cell. The low-abundance PBGD transcript was readily detected.
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