This product provides a reliable solution for DNA isolation from yeast samples. Total DNA can be purified from yeast (<5x 107) without phenol or chloroform. The whole extraction can be finished within 60 minutes. Purified DNA can be directly used for PCR, Southern blot, ect.
Specifications
| Features | Specifications |
| Main Functions | Isolation total DNA from yeast cultures |
| Applications | PCR, southern blot and enzyme digestion, etc. |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Yeast culture |
| Sample amount | Bacterial culture: 1-1.5 ml |
| Elution volume | ≥30μl |
| Time per run | ≤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 | D314702 | D314703 |
| Purification Times | 50 Preps | 250 Preps |
| Hipure DNA Mini Columns I | 50 | 250 |
| 2ml Collection Tubes | 50 | 250 |
| Glass Beads (0.4~0.6mm) | 20 g | 90 g |
| Buffer SE | 30 ml | 150 ml |
| Lyticase | 1.8 ml | 5 x 1.8 ml |
| Buffer ATL | 30 ml | 150 ml |
| ReagentDX | 500 μl | 1500 μl |
| Buffer DL | 30 ml | 150 ml |
| Buffer GW1* | 13 ml | 66 ml |
| Buffer GW2* | 20 ml | 2 x 50 ml |
| Proteinase K | 12 mg | 60 mg |
| Protease Dissolve Buffer | 1.8 ml | 5 ml |
| Buffer AE | 15 ml | 30 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 dry at room temperature (15–25°C) and are stable for at least 18 months under these conditions. Buffer ATL may precipitate at low temperature. Dissolve it by 37℃ water bath.
This product provides a reliable solution for DNA isolation from yeast samples. Total DNA can be purified from yeast (
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.
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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 |
