What applications are the Watchmaker RNA Library Prep Kit suitable for?
The Watchmaker RNA Library Prep workflow is ideally suited for:
- Targeted sequencing protocols employing hybridization capture
- Whole transcriptome sequencing, where both protein coding on noncoding transcripts are interrogated, from both high-quality and degraded samples
- mRNA sequencing, when using previously enriched poly(A)-tailed RNA
- Gene expression analysis
- Isoform/splice variant/gene fusion identification
- Single nucleotide variant detection
- Novel transcript discovery
- Workflows that employ unique molecular identifiers (UMIs) for improved sensitivity
Does the Watchmaker RNA Library Prep Kit provide stranded orientation information?
Yes, the workflow provides stranded RNA-Seq libraries.
Which species are the Watchmaker RNA Library Prep Kits compatible with?
The Watchmaker RNA Library Prep Kit is compatible with total RNA from any species.
What is the starting material I need to use when preparing libraries using the Watchmaker RNA Library Prep Kit?
High-quality total RNA ranging from 0.25 – 100 ng has been tested and shown to produce high-performing libraries. Performance with particularly degraded and challenging samples can often be salvaged by increasing the total RNA input amount, with a maximum of 100 ng.
Which adapters is the Watchmaker RNA Library Prep Kit compatible with?
This kit is compatible with adapters that have a 3′ overhanging T to facilitate adapter orientation during dscDNA ligation. Note that adapter quality impacts overall library preparation efficiency. Ensure that adapters are adequately duplexed and at the appropriate concentration prior to use. For a list of commercially available adapters and more detailed adapter recommendations, please contact support@watchmakergenomics.com.
What are the RNA purity requirements?
RNA inputs should be free from contaminating DNA. If the total RNA contains DNA, remove the contamination by incubating with DNase I (not supplied with kit). RNA should be suspended in RNase-free water and be free of salts (e.g., Mg2+, or guanidinium salts), chelating agents (e.g., EDTA or EGTA), and organics (e.g., phenol or ethanol).
Is vortex mixing safe for reactions containing RNA and/or enzyme?
Vortex mixing is recommended for master mix generation and subsequent addition to sample. Pipette mixing is an acceptable alternative so long as care is taken to ensure a completely homogeneous reaction.
What is the shelf life and recommended storage conditions?
The Watchmaker RNA Library Prep Kit has a shelf life of 12 months upon receipt, provided it is stored at -20°C ±5°C and handled under recommended storage conditions.