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.
This product is suitable for rapid extraction of total DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.
Details
Specifications
| Features | Specifications |
| Main Functions | Isolation total DNA from tissue / blood / body fluid / swab /dry blood spots |
| Applications | PCR, qPCR, southern bolt and virus detection, etc |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Tissue, cell, blood, saliva, swab, blood spot,semen and other clinical samples |
| Sample amount | Solid tissue: 1-10mg; Anticoagulant blood: 200μl |
| Elution volume | ≥20μl |
| Time per run | 30 – 60 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100µg |
This product is based on silica Column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, Nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).
Kit Contents
| Contents | D301802 | D301803 |
| Purification Times | 50 | 250 |
| HiPure DNA Mini Columns I | 50 | 250 |
| 2ml Collection Tubes | 100 | 500 |
| Buffer ATL | 30 ml | 150 ml |
| Buffer AL | 30 ml | 150 ml |
| Buffer GW1 | 22 ml | 88 ml |
| Buffer GW2 | 12 ml | 50 ml |
| Proteinase K | 24 mg | 120 mg |
| Protease Dissolve Buffer | 1.8 ml | 10 ml |
| Buffer AE | 15 ml | 60 ml |
Storage and Stability
Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.
Experiment Data
This product is suitable for rapid extraction of total DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.
Giardiasis is a disease of the small bowel caused by the protozoan parasite Giardia intestinalis (syn.duodenalis or lamblia). Giardia is one of the most common intestinal parasites in the world, and occurs at very high prevalence rates in places with poor water sanitation. Individuals become infected through ingesting or coming into contact with contaminated water, food or soil. It can also be spread through the faecal-oral route due to poor hygiene practices, which makes it common in day-care centers. G. intestinalis lives inside the intestines of infected humans or other animals including cats, dogs, birds, cows, beaver and deer. Symptoms of infection include diarrhea, malaise, excessive gas, bloating, nausea, diminished interest in food, possible vomiting and weight loss.
There are 2 kits available for the detection of Giardia intestinalis:
Giardia intestinalis TaqMan RT-PCR Kit, 100 reactions
Giardia intestinalis 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. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.
| Component | Cat. TM43850 (100 preps) | Cat. TM43810 (100 preps) |
|---|---|---|
| MDx TaqMan 2X PCR Master Mix | 2 x 700 μL | – |
| Giardia intestinalis Primer & Probe Mix | 280 μL | 280 μL |
| Giardia intestinalis Positive Control | 150 μL | 150 μL |
| Nuclease-Free Water (Negative Control) | 1.25 mL | 1.25 mL |
| Product Insert | 1 | 1 |
The amount of RNA that can be extracted from different biological or clinical samples varies greatly. For example, while a few micrograms of RNA could be easily purified from tissues and cells in excess amounts (such as from a few milligrams of tissue), many liquid biopsy samples may yield very low amounts of RNA. In fact, samples such as urine or plasma may yield 1 – 100 ng or less RNA per 100 µL of sample. Such a range of RNA quantity is often below the detection limit of most commonly used techniques for measuring RNA including nano-spectrophotometry and fluorescent nucleic acid stains. As a result, without properly determined RNA concentration, it becomes very difficult to normalize the starting quantity of RNA used in gene expression studies.
Norgen’s microRNA (cel-miR-39) Spike-In Kit offers a quantified synthetic RNA (cel-miR-39) for spike-in during RNA extraction procedures and subsequent normalization in RT-qPCR assays. The amount of cel-miR-39 RNA recovered after RNA extraction is directly correlated with the amount of total RNA recovered. After reverse transcription (such as with Norgen’s microScript Reverse Transcription system) of the sample RNA (with spike-in), the level of cel-miR-39 can be determined by subjecting the cDNA generated to quantitative PCR (qPCR) using fluorescent nucleic acid stains such as SYBR Green. A cel-miR-39 specific primer is included in the kit. The level of expression of any target transcripts in different RNA samples can now be normalized to the cel-miR-39 transcript level using standard method such as ∆∆Ct relative quantification.
In addition, the cel-miR-39 RNA is compatible to library preparation methods (including ligation-based protocols) in Next Generation Sequencing (Small RNA-Seq) workflows. The cel-miR-39 RNA could be used for normalization as well as for tracking library construction efficiency.
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Storage Conditions
Upon receipt, store Norgen’s microRNA (cel-miR-39) Spike-In Kit at -20°C or lower. Avoid multiple freeze-thaw cycles. If needed, prepare smaller working aliquots and store at -20°C or lower.
| Component | Cat. 59000 |
|---|---|
| cel-miR-39 RNA | 10 pmol |
| cel-miR-39 Forward PCR Primer | 1 nmol |
| Nuclease-Free Water | 1.25 mL |
| Product Insert | 1 |
