MessageBOOSTER™ cDNA Synthesis from Cell Lysates Kit
Produce amplified amounts of cDNA from precious, limiting cell samples without introducing 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.
- 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.
The MessageBOOSTER™ cDNA Synthesis from Cell Lysates Kit enables the user to perform sensitive qRT-PCR studies from very small populations of cells, even from as little as one cell.
The kit amplifies the cellular mRNA directly from a cell lysate, without the need to purify total RNA, and then converts the amplified RNA to cDNA that is ready for qPCR (Fig. 1). Amplified cDNA is efficiently produced from the lysates of 1 to 500 cells.
| ||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. |
Contents: QuickExtract™ RNA Extraction Solution, MessageBOOSTER™ T7-Oligo(dT) Primer, MessageBOOSTER™ Reverse Transcription PreMix, RNase Inhibitor, MMLV Reverse Transcriptase, MessageBOOSTER™ DNA Polymerase PreMix, MessageBOOSTER™ DNA Polymerase, MessageBOOSTER™ cDNA Finishing Solution, MessageBOOSTER™ In Vitro Transcription PreMix, MessageBOOSTER™ T7 RNA Polymerase, MessageBOOSTER™ T7 Transcription Buffer, RNase-Free DNase I, Random Primers, RNase H, Forward Control PCR Primer, Reverse Control PCR Primer, 100 mM DTT, RNase-Free Water, Poly(I).