3rd July 2024

TargetAmp™ 1-Round Biotin-aRNA Amplification Kit 105

TargetAmp™: Selection Guide | Process | Performance | FAQ | Poster (3 MB PDF)

Applications

  • Amplification and biotin-labeling of mRNA in total RNA of >2,500 cells (≥25 ng RNA) for use in microarray analysis.

The TargetAmp™ 1-Round Biotin-aRNA Amplification Kit 105* produces microgram amounts of biotin-antisense RNA (biotin-aRNA, also called biotin-cRNA) from as little as 25 ng of total cellular RNA. The kit provides all reagents for the preparation of biotin-aRNA (except reverse transcriptase and RNA purification columns) and uses an improved Eberwine-type linear amplification method1 to achieve at least 5,000-fold amplification of the poly(A) RNA present in a total cellular RNA sample. The entire procedure can be completed in 1 day.

Benefits

  • Simple, single-tube, 6-hour procedure. Prepare biotin-aRNA and begin hybridization the same day.
  • Biotin-aRNA produced by the kit detected more transcripts (genes) than competitive kits and showed high correlation with MAQC TaqMan® data (Fig. 1).
  • Multiple RNA samples can be easily amplified and labeled simultaneously.
  • The kit generates microgram amounts of biotin-aRNA from as little as 25 ng of input total cellular RNA (Table 1).
  • Linear RNA amplification process preserves the transcript profile of the sample.
  • Highly reproducible amplification, labeling, and microarray results.

References

  1. Van Gelder, R.N. et al. (1990) Proc. Natl. Acad. Sci. USA 87, 1663.

 
Figure 1. Correlation of expression ratios obtained from qPCR and from microarrays hybridized to aRNA prepared with the TargetAmp™ 105 kit. X axis, log2 expression ratios determined from Human Genome U133 Plus 2.0 Arrays (Affymetrix) hybridized to targets prepared by the TargetAmp 105 kit; Y axis, log2 expression ratios determined by the MAQC, using TaqMan® assays. The high correlation obtained (r = 0.936) indicates the excellent fidelity of the amplification process.

 

Table 1. A TargetAmp™ Kit 105 reaction produces high yields of biotin-aRNA from as little as 25 ng of input total RNA. Results represent the average of multiple experiments.

Amount of Input HeLa Total RNA Biotin-aRNA Yield from HeLa Total RNAa
25 ng 4.1 µg
100 ng 13.7 µg
500 ng 64.9 µg

a. Approximately 2% of HeLa total RNA is poly(A) RNA.

*TargetAmp™ aRNA Amplification Products are covered by intellectual property licensed to Epicentre Technologies Corporation from Johnson & Johnson Pharmaceutical Research & Development, L.L.C., and by intellectual property sublicensed to Epicentre Technologies Corporation from Incyte Corporation.

For research use only. Some applications in which this product may be used may be covered by patents or patent applications applicable in certain countries. Because purchase of this product does not include a license to perform any patented application, users of this product may be required to obtain a license, depending upon the particular application and country in which the product is used.

 

Ordering

Catalog No. Size
TargetAmp™ 1-Round Biotin-aRNA Amplification Kit 105
TAB1R80524 24 Reactions
These kits were optimized using SuperScript™ III reverse transcriptase (Invitrogen), which is not included in the kits. Contents: TargetAmp™ T7-Oligo(dT) Primer A, TargetAmp™ Reverse Transcription PreMix, TargetAmp™ DNA Polymerase PreMix I, TargetAmp™ DNA Polymerase, TargetAmp™ T7 RNA Polymerase, TargetAmp™ T7 Transcription Buffer, TargetAmp™ NTP PreMix, TargetAmp™ UTP / Biotin-UTP, RNase-Free DNase I, DTT, HeLa Total RNA Control, RNase-Free Water.