TIGIT is an immune receptor present on some T cells and Natural Killer cells. TIGIT binds with high affinity to the poliovirus receptor (PVR) which causes increased secretion of IL10 and decreased secretion of IL12B and suppresses T cell activation by promoting the generation of mature immunoregulatory dendritic cells. Through the CD226/TIGIT-PVR pathway, TIGIT regulates T cell mediated immunity. In cancer, TIGIT and PD-1 have been shown to be over-expressed on tumor antigen-specific CD8+ T cells and CD8+ tumor infiltrating lymphocytes (TILs) from individuals with melanoma. Blockade of TIGIT and PD-1 led to increased cell proliferation, cytokine production, and degranulation of tumour antigen-specific CD8+ T cells and TIL CD8+ T cells. It can be considered an immune checkpoint.
The HiPure Liquid RNA & miRNA Kit integrates phenol/guanidine-based sample lysis and silica membrane purification of total RNA. MagZol LS Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate designed to facilitate lysis of liquid sample and inhibit RNase. The high lysis efficiency of the reagent and the subsequent removal of contaminants by organic phase extraction enable the use of up to 0.25ml liquid sample per mini spin column.
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA from 0.25ml body fluids using column and MagZol LS reagent |
| Applications | qPCR / RT-PCR, liquid or solid-phasechip analysis, hybridization and SNP detection |
| Purification method | Mini spin column |
| Purification technology | Silica technology, acidphenol / guanidine extraction technology (MagZol pretreatment technology) |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Blood, serum, plasma |
| Sample amount | 250μl |
| Elution volume | ≥20μl |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100μg |
0.25ml liquid samples are homogenized in 0.75ml MagZol LS Reagent. After addition of chloroform, the homogenate is separated into aqueous and organic phases by centrifugation. The upper, aqueous phase is extracted, and ethanol is added to provide appropriate binding conditions. The sample is then applied to the spin column, where the total RNA (up to 100 µg) binds to the membrane and phenol and other contaminants are efficiently washed away. High-quality RNA is then eluted in 30–100µl of RNase-free water.
Advantages
Kit Contents
| Contents | R416302 | R416303 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure RNA Mini Columns | 50 | 250 |
| 2ml Collection Tubes | 50 | 2 x 125 |
| MagZol LS Reagent | 60 ml | 270 ml |
| Buffer RWC | 20 ml | 60 ml |
| Buffer RW2* | 20 ml | 2 x 50 ml |
| RNase Free Water | 10 ml | 30 ml |
Storage and Stability
MagZol LS Reagent should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions.
The HiPure Liquid RNA & miRNA Kit integrates phenol/guanidine-based sample lysis and silica membrane purification of total RNA. MagZol LS Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate designed to facilitate lysis of liquid sample and inhibit RNase. The high lysis efficiency of the reagent and the subsequent removal of contaminants by organic phase extraction enable the use of up to 0.25ml liquid sample per mini spin column.
Norovirus is a single-stranded, non-enveloped RNA virus belonging to the Caliciviridae family. Norovirus is considered the major causative agent of non-bacterial gastroenteritis in the world. The main channel of transmittance is via contaminated food or water, as well as human-to-human contact. Most Noroviruses infecting humans belong to genogroup I and II (particularly genotype 4). Norovirus is very stable in the environment and is resistant to some surface disinfectants such as alcohols and detergents. Moreover, it is very difficult to culture Norovirus in vitro, making its identification by standard microbiological assays challenging.
NoV TaqMan RT-PCR Kit, 100 reactions
NoV TaqMan RT-PCR Probe/Primer Set and Controls, 100 reactions
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Storage Conditions and Product Stability
All kit components can be stored for 1 year after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival.
| Component | Cat. TM41450 (100 preps) | Cat. TM41410 (100 preps) |
|---|---|---|
| MDx TaqMan 2X PCR Master Mix | 2 x 700 μL | – |
| NoV Primer & Probe Mix | 280 μL | 280 μL |
| NoV Positive Control | 150 μL | 150 μL |
| Nuclease-Free Water (Negative Control) | 1.25 mL | 1.25 mL |
| Product Insert | 1 | 1 |
