Description
Specifications
| Clone | IHC410 |
| Source | Mouse Monoclonal |
| Positive Control | Colon, Colon Carcinoma |
| Dilution Range | 1:200 |
MutS Homolog 2 (MSH2) is a protein involved in the mismatch-repair pathway. This protein is commonly associated with hereditary non-polyposis colorectal cancer, and mutations in this gene are correlated with the development of sporadic colorectal carcinoma. Expression levels of MSH2 are abnormally low in a high percentage of patients with microsatellite instability, as well as endometrial and ovarian cancers. Use of Anti-MSH2 is optimized when paired in an IHC panel with antibodies against MSH6, MLH1, and PMS2. Reports have shown Anti-MSH2 to be useful in the detection of the protein in a number of normal and neoplastic tissues, and for identifying a loss of MSH2 in tumors that are microsatellite-unstable.
| Clone | IHC410 |
| Source | Mouse Monoclonal |
| Positive Control | Colon, Colon Carcinoma |
| Dilution Range | 1:200 |
Sample type purification kit guide
The 16S V1-V2 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V1-V2 amplicon libraries to be used for next-generation sequencing on Illumina platforms. All molecular reagents including primers, enzyme mixes, indexes, and buffers are provided. Instructions for PCR clean up with the AMPure XP Magnetic Beads (supplied by customer) are also included for rapid purification of nucleic acid products generated at two steps of the workflow. The library prep workflow could be used for purified DNA inputs from different sources including stool, soil, water, saliva, plant, urine, skin swab, vaginal swab, cheek swab, nasal swab, plasma/serum, tongue swab, gum swab, and others.
The 16S V1-V2 Library Preparation Kit for Illumina has a streamlined procedure that reduces the handling time such that the library prep procedure can be completed in approximately 4 hours (see diagram below). Input DNA is first subjected to targeted PCR to amplify the V1-V2 region of the DNA encoding 16S rRNA. The post-PCR reaction is then cleaned up using AMPure XP beads. Dual index primers are then added using a limited-cycle PCR. The indexed amplicons flanked by 5′ and 3′ barcoded adaptors are then cleaned using AMPure XP beads. The libraries are then ready for quantification, pooling and sequencing.
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| Minimum amount of starting material: | 2.5 µL of DNA (5 ng/µL) |
| Time to complete library preparation: | 4 hours |
Storage Conditions and Product Stability
Norgen’s 16S V1-V2 Library Prep Kit for Illumina is shipped as one kit box (for the 24 prep kit) or two sub-component kits (for the 96 prep kit). All kits should be stored at -20°C upon arrival.
All kit components should remain stable for at least 1 year when stored at the specified storage conditions.
| Step | Component | Cat. 70100 (24 preps) | Cat. 70110, 70120, 70130, 70140 (96 preps) |
|---|---|---|---|
| Amplicon PCR (PCR 1) | MGX Master Mix | 330 µL | 1,320 µL |
| 16S V1-V2 Primer Mix | 70 µL | 280 µL | |
| Index PCR (PCR 2) | Indexing Master Mix | 660 µL | 2 x 1,320 µL |
| N7xx Index Primer | 50 µL | 50 µL | |
| S5xx Index Primer | 70 µL | 70 µL | |
| PCR Clean-Up | Resuspension Buffer | 2 x 1,250 µL | 2 x 5,000 µL |
| Nuclease-free water | 1,250 µL | 1 x 6,000 µL |
These kits provide a fast, reliable and convenient spin column method for the isolation of high quality, high purity and inhibitor-free cell-free circulating DNA (cfc-DNA) from plasma/serum sample. These kits are designed to isolate all sizes of cfc-DNA from either fresh or frozen plasma/serum samples and the purified DNA is eluted into a flexible elution volume ranging from 25 µL to 50 µL. The purified plasma/serum cfc-DNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive PCR and Southern Blot analysis, microarrays and NGS.
Background
Plasma/Serum cell-free circulating DNA (cfc-DNA) has the potential to provide biomarkers for certain cancers and disease states as well as fetal DNA in maternal blood. Currently, significant advancements are being made in utilizing cfc-DNA as biomarkers for the early diagnosis, prognosis and monitoring of therapy for several cancer types and autoimmune diseases. Cell-free mitocondrial DNA (cfmtDNA) is also under investigation for its clinical significance. This cfc-DNA is usually present as short fragments of less than 1000 bp. In addition, cell-free fetal DNA has been widely used as a non-invasive method for prenatal diagnosis including early identification of fetal sex, genetic studies for families at high risk for inherited genetic disorders, screening for Rhesus factor, screening for aneuploidy and identification of preeclampsia.
Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit Dx
For serum input volumes 10 µL – 200 µL. Purify high-quality DNA in 15-20 minutes
Plasma/Serum Cell-Free Circulating DNA Purification Mini Kit Dx
For serum input volumes 200 µL – 500 µL. Purify high-quality DNA in 15-20 minutes.
Plasma/Serum Cell-Free Circulating DNA Purification Midi Kit Dx
The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 25 µL to 50 µL. All components for the purification & concentration are provided in one convenient & fast kit for the easy processing of large input volumes of bodily fluids. For a schematic workflow of the protocol click here to view. For serum input volumes 1 mL – 4 mL. Purify high-quality DNA in 90 minutes.
Plasma/Serum Cell-Free Circulating DNA Purification Maxi Kit Dx
The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 25 µL to 50 µL. All components for the purification & concentration are provided in one convenient & fast kit for the easy processing of large input volumes of bodily fluids. For a schematic workflow of the protocol click here to view. For serum input volumes 5 mL – 10 mL. Two kits in one – purify and concentrate DNA from bodily fluids using a convenient two column system.
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| Kit Specifications | |
| Minimum Plasma/Serum Input | 10 μL |
| Maximum Plasma/Serum Input | 200 μL |
| Size of DNA Purified | ≥ 50 bp |
| Elution Volume | 25-50 μL |
| Time to Complete 10 Purifications | 15-20 minutes |
| Average Yields* | Variable depending on specimen |
*Please check page 7 in the product manual for the Plasma/Serum Average Yields and the Common DNA Quantification Methods.
Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature (15-25°C) for up to 2 years without showing any reduction in performance. Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Kits contain ready-to-use Proteinase K solution, which is dissolved in a specially prepared storage buffer. The Proteinase K is stable for up to 2.5 years after delivery when stored at room temperature. To prolong the lifetime of Proteinase K, storage at 2–8°C is recommended.
| Component | Cat. Dx55100 (50 preps) | Cat. Dx55500 (50 preps) | Cat. Dx55600 (20 preps) | Cat. Dx55800 (10 preps) |
|---|---|---|---|---|
| Binding Buffer B | 2 x 40 mL | 40 mL | 2 x 85 mL 1 x 12 mL | 2 x 85 mL 1 x 40 mL |
| Proteinase K | 2 x 1 mL | 0.6 mL | 6.5 mL | 8 mL |
| Solution WN | 18 mL | 18 mL | 18 mL | 18 mL |
| Wash Solution A | 38 mL | 18 mL | 18 mL | 18 mL |
| Elution Buffer B | 8 mL | 8 mL | 30 mL | 30 mL |
| Micro Spin Columns | 50 | 50 | 20 | 10 |
| Mini Spin Columns | 50 | – | – | – |
| Midi Spin Columns | – | – | 20 | – |
| Maxi Spin Columns | – | – | – | 10 |
| Collection Tubes | 100 | 50 | 20 | 10 |
| Elution Tubes (1.7 mL) | 100 | 50 | 20 | 10 |
| Product Insert | 1 | 1 | 1 | 1 |
HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA from FFPE tissue and section samples (with DNase) |
| Applications | RT-PCR, quantitative RT-PCR, Northern hybridization, Poly A purification, nucleic acid protection and in vitro translation |
| Purification method | Mini spin column |
| Purification technology | Silica technology, DNase |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | FFPE tissue sample |
| Sample amount | 6mg |
| Yield | 20μg |
| Elution volume | ≥10μl |
| Time per run | ≤60 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100µg |
This product is based on silica column purification. Remove paraffin by Buffer DPS. Sample lysis with proteinase K digestion requires only 15 minutes. After lysis, samples are incubated at 80ºC for 15 minutes. Transfer to an adsorption column and RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, RNA was finally eluted with low-salt buffer.
Advantages
Kit Contents
| Contents | R414402 | D414403 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure RNA Micro Columns | 50 | 250 |
| 2ml Collection Tubes | 50 | 250 |
| Buffer DPS | 60 ml | 250 ml |
| Buffer FRL | 15 ml | 60 ml |
| Buffer RLC | 15 ml | 60 ml |
| Buffer RWC* | 10 ml | 50 ml |
| Buffer RW2* | 20 ml | 2 x 50 ml |
| DNase I | 600 µl | 5 x 600 µl |
| DNase Booster Buffer | 1.5 ml | 6 ml |
| Protease Dissolve Buffer | 1.8 ml | 10 ml |
| Proteinase K | 24 mg | 120 mg |
| RNase Free Water | 10 ml | 20 ml |
Storage and Stability
Proteinase K should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K up to 8 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.
Experiment Data
HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.