What applications is the Watchmaker mRNA Library Prep Kit suitable for?
- Gene expression analysis
- Gene fusion detection
- Isoform/alternative splicing analysis
- Novel transcript discovery
- Single nucleotide variant (SNV) detection
- Pathway analysis
Which species are the Watchmaker mRNA Library Prep Kits compatible with?
The Watchmaker mRNA Library Prep Kit is compatible with eukaryotic species.
Why use mRNA enrichment?
mRNA enrichment focusses sequencing on coding transcripts. If your application only requires coverage of coding transcripts, then mRNA enrichment provides improved sequencing economy by focusing on transcripts you care about.
Does the Watchmaker mRNA Library Prep Kit provide strand orientation information?
Yes, the workflow provides stranded RNA-Seq libraries.
Does this product support long-read sequencing?
We use random primers to prime during first strand synthesis, which does not support full-length cDNA generation. This makes it unlikely to be compatible with long-read applications.
What sample types are compatible with the Watchmaker mRNA Library Prep Kit?
Intact (RIN >7) RNA ranging from 2.5 – 1000 ng is compatible with the Watchmaker mRNA Library Prep Kit. This includes RNA derived from tissues and cell lines, suspended in 50 µl of RNase-free water.
This kit is not compatible with degraded samples, such as FFPE. To prepare libraries from degraded inputs, please use the Watchmaker RNA Library Prep Kit with Polaris Depletion.
For inputs outside this range, 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).
Can this kit be used to detect non-coding RNAs?
This kit enriches for coding RNA. To detect non-coding RNAs, please use the Watchmaker RNA Library Prep Kit with Polaris Depletion.
Which adapters are the Watchmaker mRNA 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.
Is this kit compatible with other (non-Illumina) sequencing platforms?
This kit has been demonstrated to be compatible with Element, Singular and Ultima sequencing platforms.
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 mRNA Library Prep Kit has a shelf life of 6 months upon receipt. The mRNA capture Kit should be stored at 2 – 8°C and the RNA Library Prep Kit at -25°C to -15°C. Both kits should be handled under recommended conditions.
Which modules are included in the Watchmaker mRNA Library Prep Kit? ?
The mRNA Library Prep Kit (7BK0001) consists of the mRNA capture kit (7K0105) and RNA Library Prep Kit (7K0078). You will receive two boxes, labeled with part numbers 7K0105 and 7K0078.
What is the purpose of the FFPE Treatment Buffer included in the RNA Library Prep module?
The FFPE treatment buffer in the RNA Library Prep Kit box is not used as part of the mRNA Library Prep Kit workflow. This tube may be removed from the kit and discarded.
What other RNA Library Prep Kits does Watchmaker offer?
The Watchmaker RNA Library Prep Kit with Polaris Depletion removes uninformative ribosomal and globin RNAs allowing for whole transcriptome analysis, including long noncoding RNAs. This kit is compatible with both high quality and degraded RNA samples.
The Watchmaker RNA Library Prep Kit is suitable for total RNA and typically used upstream of target enrichment. This kit is compatible with both high quality and degraded RNA samples.
Can this kit be automated?
The Watchmaker mRNA Library Prep Kit is highly automatable. The kit was developed with automation in mind and 96 reaction kits include generous overages to account for dead volumes on liquid handling platforms. Visit our automation page for a list of scripts already developed or to contact a member of our automation team regarding your platform and needs.
Do you offer custom formats?
Yes! Watchmaker offers customer fills, packaging, concentrations, and labeling – including private label – designed to meet your unique needs. We offer flexible terms to serve organizations of any size, and our right-sized processes enable rapid turnaround time on customization. Please contact sales@watchmakergenomics.com to learn more about our capabilities.
Are you ISO 13485-certified?
Yes! Our Quality Management System has achieved ISO 13485:2016 Certification. Our certificate has been awarded for the design, development, manufacture, contract manufacture, and support of high performing reagents for genomics applications in medical research. Download the certificate here.