Hello Petra,
CleanTag library preparation is a ligation based method that is designed to tag adapters onto small RNA species with a 3′ OH and 5′ P. piRNA is thought to have this type of structure and would therefore be compatible with the CleanTag kit.
Best Regards,
Joey
Both products included the reagents needed for the ligation of the 3′ adapter and 5 ‘ adapter. The CleanTag™ Small RNA Library Prep Kit (Cat# L-3206) also includes the reagents needed for the RT and PCR steps of library preparation which were not included in the CleanTag™ Ligation Kit (Cat# L-3203).
| CleanTag™ Small RNA Library Prep Kit (Cat# L-3206) |
CleanTag™ Ligation Kit (Cat# L-3203) |
| CleanTag™ 3′ Adapter | CleanTag™ 3′ Adapter |
| CleanTag™ 5′ Adapter | CleanTag™ 5′ Adapter |
| Enzymes, 1 & 2 | Enzymes, 1 & 2 |
| Buffers, 1 & 2 | Buffers, 1 & 2 |
| Reverse Transcriptase | |
| RT Buffer | |
| dNTP Mix (10 mM ea) | |
| DTT | |
| High Fidelity PCR Master Mix (2X) | |
| RNase Inhibitor | RNase Inhibitor |
The CleanTag™ Ligation Kit was discontinued in early 2016.
We recommend a two step magnetic bead purification using Agencourt AMPure® XP – PCR Purification (A63880) by Beckman Coulter. The protocol can be found on the product insert for the CleanTag™ Small RNA Library Prep Kit (Cat# L-3206).
While we recommend sticking to 2 uL of RNA input when possible, we found that we still obtain good library prep results with up to 10 uL of RNA (without changing any other volumes).
Yes.
Buffer components may cause higher base pair shift in crude Bioanalyzer® traces as show below. miRNA peak should be at ~150 bp for crude samples and at ~140 bp when purified. piRNA peak should be at ~ 160 for crude samples and at ~150 bp when purified.
Diluted Crude Sample
Purified Sample
CleanTag™ Small RNA Library Prep Kit (Cat# L-3206) contains adapters compatible with the Illumina® sequencing platform. However, you can easily convert your tagged library into Ion Torrent™ compatible sequences during the PCR step. Contact us to discuss.
The CleanTag™ Small RNA Library Prep Kit (Cat# L-3206) has been specifically formulated to work with CleanTag™ adapters. We strongly recommend using our workflow to ensure best results.