Shop All Nascent RNA Detection & Capture Kits

Click-iT™ Nascent RNA Capture Kit, for gene expression analysis (Invitrogen™)

The Click-iT® Nascent RNA Capture Kit, uses our proprietary Click-iT® chemistry to capture with high efficiency and sensitivity newly minted RNAs. By the Click-iT® approach studies on high resolution analysis of gene regulation can be performed along with RNA regulation, RNA stability, RNA synthesis, RNA decay and degradation, transcriptional regulation, delta⁄delta C(t), nuclear run on⁄run off and more. Ethylene uridine (EU) ribonucleotide homologs containing an alkyne reactive group are fed to cells or tissue. Once incorporated, cells are lysed and the RNA isolated. A biotin azide is 'clicked on’ and then strepavidin magnetic beads are used to capture the newly synthesized pool of RNA. The captured RNAs are then amplified and the resultant cDNAs are used on any number of post-capture methodologies. We have validated it for arrays (both high and low density), sequencing on SOLiD™ and either SYBR® or TaqMan® based qPCR based delta CT analysis. As few as 25,000 cells are needed for the analysis of newly induced transcripts. The kit allows for 40 conditions for labeling and capture that can then be divided into 400 separate analytical assayss. With a price of around dollar an assy this is perfect for researchers in academia and industrial settings.

The EU reagent at the recommend concentration of 200 µM is well tolerated by cells. In an array screen of over 32,000 genes we could detect only the most minor changes in 12 compared to a control vehicle. Furthermore at initial feeding concentration 5x of what we recommend (200 µM vs 1.0 mM) there were no effects on a panel of housekeeping genes, or on the cells ability to proliferate normally. Sensitivity and reliability was confirmed in a low density array (TLDA®) of apoptotic genes. The captured nascent pool accurately found all the previously characterized apoptotic (stauroporine induced) transcripts, without any detectable change in sequence information.

*Discovery not just recovery.* The last decades Genomic efforts revealed that only 2% of the human encodes protein, leaving 98% of the human genome as the Unknome. Discovering the purpose and expression patterns of this vast reservoir of unknown sequence will be markedly aided in this decade’s transcriptomics by this new approach.

*Ability to study RNA Stability. * For others the ability to discover the stability of mRNA’s, without the need for radioactivity will be the key feature offered here. In the past transcript half lives were derived with radioactive nucleotides in traditional nuclear run and run off experiments. Cumbersome and dangerous, this approach is seen a steep decline – now these critical values can be derived simply, safely and reliably.

*Want to know what’s new?* Incorporate the Click-iT Line of metabolic analogs to label the new stuff! Without radioactivity or difficult to use haptens. Find what’s new in DNA, RNA, protein, sugars, ptms and more.
*Click-iT® Technology - One Reaction, Endless Possibilities!*

Click-iT™ RNA Alexa Fluor™ 594 Imaging Kit (Invitrogen™)

The Click-iT® RNA Alexa Fluor® 594 Imaging Kit enables detection of global RNA transcription temporally and spatially in cells and tissues (PNAS (2008) 105:15779-84).The ability to detect newly synthesized RNA or changes in RNA levels resulting from disease, environmental damage or drug treatments is an important aspect of toxicological profiling. Utilizing an alkyne-modified nucleoside, 5-ethynyl uridine (EU), and powerful click chemistry, newly synthesized RNA can be detected without the use of radioactivity or antibodies with a simple, two-step procedure. In step one, the alkyne-containing nucleoside is fed to cells or animals and actively incorporated into nascent RNA. The small size of the tag enables efficient incorporation of the modified nucleoside into RNA, but not into DNA. Detection utilizes the chemoselective ligation or “click" reaction between and azide and an alkyne where the modified RNA is detected with a corresponding azide-containing dye. With its dimunitive “footprint", the Click-iT® detection molecule can easily penetrate complex samples and leaves open the possibility of multiplex analyses with other probes, including antibodies for the detection of RNA-interactive proteins for deeper biological insights.

Click-iT™ RNA Alexa Fluor™ 488 Imaging Kit (Invitrogen™)

The Click-iT® RNA Alexa Fluor® 488 Imaging Kit enables detection of global RNA transcription temporally and spatially in cells and tissues (PNAS (2008) 105:15779-84).
The ability to detect newly synthesized RNA or changes in RNA levels resulting from disease, environmental damage or drug treatments is an important aspect of toxicological profiling. Utilizing an alkyne-modified nucleoside, 5-ethynyl uridine (EU), and powerful click chemistry, newly synthesized RNA can be detected without the use of radioactivity or antibodies with a simple, two-step procedure. In step one, the alkyne-containing nucleoside is fed to cells or animals and actively incorporated into nascent RNA. The small size of the tag enables efficient incorporation of the modified nucleoside into RNA, but not into DNA. Detection utilizes the chemoselective ligation or “click" reaction between and azide and an alkyne where the modified RNA is detected with a corresponding azide-containing dye. With its dimunitive “footprint", the Click-iT® detection molecule can easily penetrate complex samples and leaves open the possibility of multiplex analyses with other probes, including antibodies for the detection of RNA-interactive proteins for deeper biological insights.

Click-iT™ RNA Alexa Fluor™ 488 HCS Assay (Invitrogen™)

The Click-iT® RNA Alexa Fluor® 488 HCS Assay enables detection of global RNA transcription temporally and spatially in cells (PNAS (2008) 105:15779-84). The ability to detect newly synthesized RNA or changes in RNA levels resulting from disease, environmental damage or drug treatments is an important aspect of toxicological profiling. Utilizing an alkyne-modified nucleoside, 5-ethynyl uridine (EU), and powerful click chemistry, newly synthesized RNA can be detected without the use of radioactivity or antibodies with a simple, two-step procedure. In step one, the alkyne-containing nucleoside is fed to cells or animals and actively incorporated into nascent RNA. The small size of the tag enables efficient incorporation of the modified nucleoside into RNA, but not into DNA. Detection utilizes the chemoselective ligation or “click" reaction between and azide and an alkyne where the modified RNA is detected with a corresponding azide-containing dye. With its dimunitive “footprint", the Click-iT® detection molecule can easily penetrate complex samples and leaves open the possibility of multiplex analyses with other probes, including antibodies for the detection of RNA-interactive proteins for deeper biological insights.