Figure 1 / 1
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Ladder Properties:
• 22 discrete bands, ranging from 60 bp to 600 bp
Storage:
• Store at -20°C in small aliquots. For longer term storage, -70°C is recommended.
• The product is light-sensitive. Minimize light exposure.
This kit provides fast purification of high-quality RNA from cells, tissues, and yeast using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol/chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. RNA purified using the HiPure Total RNA Purification System can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
Specifications
| Features | Specifications |
| Main Functions | Isolation total RNA (not include miRNA) from 20mg tissue, 150mg plant, 5 x 106 cell using two columns (gDNA removed column) |
| Applications | RT-PCR, qRT-PCR, Northern hybridization, second generation sequencing |
| Purification method | Mini spin column |
| Purification technology | Silica technology, DNA filtration technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Animal soft tissue, cultured cells, lymphocytes, simple plant tissue |
| Sample amount | Cells:≤1 x 107 Animal tissue sample: 1-20 mgPlant tissue: 50-150 mg |
| Yield | 2-100μg |
| Elution volume | ≥50μl |
| Time per run | ≤25 minutes（1-24 samples） |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100µg |
The Kit isolates total RNA from up to 107 cells or 20 mg tissue. A short workflow enables RNA isolation with genomic DNA removal in less than 25 mins. Samples are first lysed and homogenized. The lysate is passed through a DNA Mini column, ethanol is added to the flow-through, and the sample is applied to a RNA column. RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in as little as 30µl water using the Kit.
Advantages
Kit Contents
| Contents | R411102 | D411103 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure DNA Mini Columns | 50 | 250 |
| HiPure RNA Mini Columns | 50 | 250 |
| 2ml Collection Tubes | 100 | 2 x 250 |
| Buffer RLC | 50 ml | 200 ml |
| Buffer RW1 | 50 ml | 200 ml |
| Buffer RW2* | 12 ml | 2 x 50 ml |
| RNase Free Water | 10 ml | 30 ml |
Storage and Stability
HiPureKit can be stored dry at room temperature (15-25°C) and are stable for at least18 months under these conditions. During shipment, crystals or precipitationmay form in the Buffer RLC. Dissolve by warming buffer to 37°C.
This kit provides fast purification of high-quality RNA from cells, tissues, and yeast using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol/chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. RNA purified using the HiPure Total RNA Purification System can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
Sample type purification kit guide
The 16S V2-V3 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V2-V3 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 V2-V3 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 V2-V3 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.
Figure 1 / 3
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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 V2-V3 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. 70300 (24 preps) | Cat. 70310, 70320, 70330, 70340 (96 preps) |
|---|---|---|---|
| Amplicon PCR (PCR 1) | MGX Master Mix | 330 µL | 1,320 µL |
| 16S V2-V3 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 |