Are you working with RNA and focusing especially on miRNAs? We at Immuno have great solutions for your needs:
Extract your RNA with Lexogen SPLIT
The SPLIT RNA Extraction Kit enables a fast (30min) and highly efficient extraction of RNA that is free of genomic DNA contamination. The RNA can be recovered as total RNA or split into two fractions, large RNA and small RNA, facilitating the analysis of e.g. mRNA and miRNA from the same sample. Thus, the RNA obtained is ideal for seamlessly preparing libraries for NGS of total RNA or its large and small fractions or any other demanding downstream application.
The SPLIT protocol provides you a splitting option with a size cut off at ~150 nt. The small RNA fraction (< 150 nt) will contain miRNA, tRNA, 5 S rRNA, and 5.8 S rRNA, while the large RNA fraction (>150 nt) will contain mRNA, 18 S rRNA, 28 S rRNA, and snRNA. Species-independent extraction! 3-prep trial kit available for testing!
Prepare your small RNA libraries with Lexogen Small RNA-Seq Library Prep Kit
Optimized solution for generating small RNA libraries for Illumina sequencing directly from total RNA or enriched small RNA. Works perfectly also with challenging low input samples – Exceptional miRNA Discovery
- Discovery and profiling of small RNA (≤ 200 nt), including miRNA, piRNA, siRNA, snoRNA, tsRNA, srRNA, U-RNA
- Wide input range from 50 pg to 1 µg of RNA
- Optimized for low RNA content samples such as liquid biopsies including exosomes
- All-in-one protocol – only 75 minutes hands-on time
- High reproducibility for inter-replicates and across concentrations
- Exceptional miRNA discovery compared to other protocols, in particular for low RNA inputs
Combine with SPLIT RNA Extraction Kit and enriched miRNA content – Complete solution for small RNA analysis!
Quantify your miRNAs with fast and sensitive Biovendor’s kits
Novel and sensitive ELISA method for miRNA quantification applicable to studies on cancer, cardiovascular and pulmonary diseases as well as regulation of immunological processes.
- RT-qPCR assay library of all miRNAs referenced in miRBase.
- Spesific: three miRNA-specific primers to discriminate single nucleotide differences
- Optimized RT-qPCR primers and reagents for efficient target amplification
- Only 1 pg of sample needed
- Wet-lab validated according to MIQE guidelines using both synthetic miRNAs and RNA from biological samples
NEW extremely specific 2-tailed assays for microRNA -exceeding performance compared to other techniques
The challenge detecting small microRNAs is that two conventional PCR primers do not fit the target as their combined length is almost twice the length of the microRNA. Older techniques have solved this by extending the microRNA using e.g., a hairpin primer, adding a poly A-tail, or adding a fragment by ligation. This, however, compromises the assay sensitivity and specificity, as only one of the PCR primers sense the actual microRNA sequence; the other senses the added extension. Further, these methods fail to detect microRNAs modified in the 3’-end as it interferes with the extension process.
TATAA miRNA two-Tailed RT-qPCR assays offer a superior solution. Instead of using a single binding probe, Two-tailed PCR uses two hemiprobes, which bind to different stretches of the microRNA, that are connected by a folded tether. While each hemiprobe is too short to bind the microRNA, when both hemiprobes are complementary they bind cooperatively.
- Binding is exceedingly specific, as a mismatch is much more profound in a short hemiprobe
- The cDNA formed can then be PCR amplified using two sequence specific primers
- SYBR used for detection
- High degree two-tube multiplexing of the RT followed by singleplex qPCR
Assays available: twelve catalog products, also Custom. Please ask for your option! Custom assays are validated on RT and qPCR level on synthetic microRNA.
Introduce your miRNAs into cells with Mirus transfection solutions
TransIT-X2® Dynamic Delivery System is the reagent of choice for delivering miRNA, miRNA mimics or pre-miRNAs.
TransIT-X2® can also be used for efficient transfection of plasmid DNA and miRNA or both.
TransIT-TKO® and TransIT-siQUEST® transfection reagents can also effectively transfect duplex miRNAs into mammalian cells in culture. Ideally, all three reagents should be tested in parallel to determine the best solution for your application. Please consult the Reagent Agent® transfection database for reference and protocols.
We are happy to tell you more!
Tuula Aaltonen, PhD, Life Science
Sanna Siltanen, PhD, NGS & Genomics
Katja-Riikka Louhi, PhD, Life Science