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
|CleanTag™ Ligation Kit
|CleanTag™ 3′ Adapter
||CleanTag™ 3′ Adapter
|CleanTag™ 5′ Adapter
||CleanTag™ 5′ Adapter
|Enzymes, 1 & 2
||Enzymes, 1 & 2
|Buffers, 1 & 2
||Buffers, 1 & 2
|dNTP Mix (10 mM ea)
|High Fidelity PCR Master Mix (2X)
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).
Thank you for your question. We recommend the Q5® High Fidelity 2x Master Mix (catalog # M0492S) from NEB for use with the CleanTag™ Ligation Kit for Small RNA Library Prep. You can view our full list of recommended reagents here.
Please let me know if you have any other questions.
You are correct in that the CleanTag™ chemistry can fully suppress adapter dimer formation. If you are using very low input levels (less than 10 ng) you may start to see some adapter dimer but it should be minimal compared to amount of tagged library. Dilution of the adapters per the product insert is key to keep adapter dimer low at lower inputs.
Yes, you can load magnetic bead purified samples without gel purification directly onto a sequencer and get good quality sequencing data.
Sabrina Shore, MS
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