MessageBooster™ Whole Transcriptome cDNA Synthesis Kit for qPCR

Catalogue number

Pack size

MBWT80510

10 reactions

The MessageBOOSTER™ cDNA Synthesis Kit for qPCR and the MessageBOOSTER™ Whole-Transcriptome cDNA Synthesis Kit for qPCR enable the user to perform hundreds of sensitive qPCR amplifications from very small populations of cells, even from as little as one cell.

Both kits amplify the mRNA contained in a small total RNA sample and then convert the amplified RNA to cDNA that is ready for PCR or qPCR.

The MessageBOOSTER™ Whole-Transcriptome cDNA Synthesis Kit for qPCR uses the proprietary MessageBOOSTER™ Whole-Transcriptome Primers to prime cDNA synthesis. These primers anneal to and prime cDNA synthesis throughout the RNA in the sample. Thus, this kit is ideal for amplifying RNA and subsequently producing cDNA from compromised or degraded RNA samples such as might be obtained by laser-capture microdissection, cell-sorting methods, or from FFPE tissue (see note 1). Note: This kit will not amplify RNA from a single cell. Use the original MessageBOOSTER™ cDNA Synthesis Kit for qPCR for single-cell amplification.

Figure 1. A MessageBOOSTER™ Whole-Transcriptome cDNA Synthesis Kit reaction amplifies the RNA from small amounts of compromised or degraded total RNA and then converts the amplified RNA to cDNA that is ready for qPCR. The single-tube reaction can be completed in 1 day.

RNA Quality	Input Total RNA	No. of qPCR Amplification
		Low-Abundance Transcripts	High-Abundance Transcripts
Partially degraded	10 ng	>10	>500
Severely degraded	100 ng	>10	>500

Table 1. The number of qPCR amplifications from cDNA produced by the MessageBOOSTER™ Whole-Transcriptome Kit.

RNA Quality	cDNA Synthesis Method	Actin (1792; high)	GAPDH (1310; high)	TFRC (5010; high)	ZNF (2878; low)	HPRT (2878; low)	TUBA (1706; low)	PDE3A (4124; medium)	ENSA (2512; low)	HMBS (1380: low)	GUSB (2245; medium)
Partially degraded	Oligo(dT)	416	64	128	5.7	9.8	832	34.3	73.5	22.6	955
Severely degraded	Oligo(dT)	1260	274	446	∞	∞	∞	∞	45.3	∞	∞
Partially degraded	MessageBooster Whole-Transcriptome Kit	2.3	0.63	1.3	1.7	1.3	12.1	1.7	3.5	1.4	4.3
Severely degraded	MessageBooster Whole-Transcriptome Kit	9.2	1.4	2.1	4.9	0.3	1.9	6.1	13.9	0.3	7.5

Table 2. Comparison of 3'/5' ratios of cDNA produced from degraded RNA samples. cDNA was produced using the MessageBOOSTER™ Whole-Transcriptome cDNA Synthesis Kit for qPCR or an oligo(dT)-primed cDNA synthesis method. The 3'/5' ratios were calculated from the qPCR CT values obtained using PCR primers pairs to the 3' or 5' end of the transcripts. A 3'/5' ratio of 1.0 demonstrates equal representation of 3' and 5' regions of the transcripts in the cDNA. A ratio of ∞ indicates that the 5' PCR amplicon was not detected.

Notes on using the MessageBOOSTER™ Whole-Transcriptome cDNA Synthesis Kit for qPCR:

1. The ability to amplify RNA from formalin-fixed paraffin-embedded (FFPE) samples using the MessageBOOSTER™ Whole-Transcriptome cDNA Synthesis Kit is dependent on factors that are often difficult for the user to control. We have observed that some RNA samples isolated from FFPE tissue have been successfully amplified and converted to cDNA while other samples have failed. Therefore, your results may vary depending on the condition of the sample.
2. Best results are obtained using degraded RNA where the average size of the RNA fragments is greater than 150 bases.
3. The amount of input total RNA per reaction can vary from 50 pg of intact RNA to 100 ng of severely degraded RNA. Guidelines are presented in the kit's literature.

Last modified: 2008-06-25 11:21:54

Applications of the MessageBOOSTER™ Whole-Transcriptome cDNA Synthesis Kit for qPCR

• Generate large amounts of cDNA from small amounts of compromised or degraded total RNA preparations for RT-PCR or qRT-PCR studies
• Generate large amounts of cDNA from small amounts of compromised or degraded total RNA preparations for archival purposes

Benefits of the MessageBOOSTER™ Whole-Transcriptome cDNA Synthesis Kit for qPCR

• Ideal for Degraded RNA. Significantly increase the number of qRT-PCR reactions that can be obtained from small amounts of degraded RNA samples
• Improved RT-PCR of Degraded RNA. Produce cDNA with greatly improved 3'/5' ratio from degraded RNA samples
• Low Input. Use as little as 1 ng of degraded total RNA per reaction
• Fast and Easy Reactions. One-day, single-tube reaction

Last modified: 2008-06-24 12:09:25

Protocols for: MessageBOOSTER™ Whole-Transcriptome cDNA Synthesis Kit for qPCR

MessageBOOSTER™ Whole-Transcriptome cDNA Synthesis Kit Protocol

(catalogue number MBWT80510)

Please note: all protocols off site are the responsiblity of the supplier

Last modified: 2008-06-24 12:06:50