Choosing RNAscope 1.0 or 2.0
RNAscope 1.0 and 2.0 kits adhere to the highest quality standards, providing ready-to-use reagent kits and probes for single-copy RNA in situ hybridization.
RNAscope 1.0 and 2.0 Reagent Kits utilize the same RNAscope Target Probe sets and are both capable of staining individual RNA molecules as distinctive dots visible under a standard bright field microscope. |
 |
How does RNAscope 2.0 differ from 1.0?
RNAscope 2.0 Reagent Kits have been specially designed to provide more intense staining for low copy target gene expression of 1-20 copies per cell to provide results that are easily visible under low power magnification (10-20x) using a standard bright field microscope. RNAscope 2.0 utilizes the same double Z probe design to ensure exceptional specificity but incorporates additional signal amplification steps to make single RNA molecules appear as larger, more intense punctate spots. In addition, RNAscope 2.0 in situ hybridization kits offer two off-the-shelf chromogenic staining options (Brown or Red).
Should I choose RNAscope 1.0 or RNAscope 2.0?
We recommend RNAscope 2.0 if you are unsure of your gene target expression level. RNAscope 2.0 can also improve results for certain applications (see Table 1 below). For example, target genes with shorter transcripts (< 800 bases) or specimens with degraded RNA (archival FFPE specimens) can yield more robust staining with RNAscope 2.0, resulting in single molecule spots that are large enough for easy visualization using low magnification under a standard bright field microscope.
Table 1. RNAscope 1.0 and 2.0 Comparison
RNAscope 1.0 |
RNAscope 2.0 |
|
Sensitivity |
Single copy |
Single copy |
Sample requirement |
FFPE |
FFPE |
Target region (typical) |
1,000 bases |
1,000 bases |
Suggested minimum target length |
800 bases |
200 bases |
Optimal expression level |
>20 copies/cell |
1-20 copies /cell |
Degraded RNA |
Less suitable |
More suitable |
Color detection |
DAB (Brown) |
DAB (Brown) or Fast Red (Red) |
Staining spot size |
Smaller |
Larger |
Staining spot observed under |
20x to 40x lens |
10x to 20x lens |
Assay time |
<7 hrs |
<8 hrs |
Housekeeping gene control |
UbC (moderate expression) |
POLR2A (low expression) |
Still not sure if you need a 1.0 or 2.0 RNAscope FFPE Reagent kit? ACD can help you determine whether a 1.0 or 2.0 reagent kit will best meet your needs. Email us at support@acdbio.com
