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 |
10 Layer Cell Factory Double Wide Mouth
Huayi Cell Factory is a robust and easy-to-use cell
culture platform for applications in the production of human and animal vaccines, therapeutic proteins, cell therapy, and gene therapy.
To address the needs of your workflow, We have three kinds of mouth, which is available on most cell culture products to ensure consistent performance from lot to lot and from format to format.
Specifications: 1 layer, 2 layers, 5 layers, 10 layers and 40 layers.
PRODUCT FEATURES
of one 10-layer Cell Factory unit is equivalent to the area
of 36 T-175 flasks
culture platform for applications in the production of human and animal vaccines, therapeutic proteins, cell therapy, and gene therapy.
To address the needs of your workflow, We have three kinds of mouth, which is available on most cell culture products to ensure consistent performance from lot to lot and from format to format.
Specifications: 1 layer, 2 layers, 5 layers, 10 layers and 40 layers.
This Kit is designed forpurification of high quality circulating DNA (cfDNA) from 1-6ml cell-free bodyfluids (such as plasma, serum). The purified DNA is suitable for direct use indownstream applications such as PCR, real-time PCR, Biochip analysis and NGS.
Specifications
Features | Specifications |
Main Functions | Isolation circulating DNA from 1-6ml plasma, serum, body fluids |
Applications | qPCR, NGS, etc. |
Purification technology | Magnetic beads technology |
Process method | Manual or automatic |
Sample type | Serum, plasma |
Sample amount | 1-6ml |
Elution volume | ≥50μl |
Time per run | ≤60 minutes |
This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and protease. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by elution buffer.
Contents | IVD5435 |
Purification Times | 50 |
MagPure Particles F | 14 ml |
Carrier RNA | 310 μg |
Proteinase K | 240 mg |
Protease Dissolve Buffer | 15 ml |
Buffer SDS | 15 ml |
Buffer MLK | 500 ml |
Buffer MAW1 | 250 ml |
Buffer MW2* | 50 ml |
Elution Buffer | 60 ml |
Storage and Stability
Proteinase K, Carrier RNA and MagPure Particles F 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.Theremaining kit components can be stored dry 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 beredissolved before use. Make sure that all buffers are at room temperature when used.
Experiment Data
This Kit is designed forpurification of high quality circulating DNA (cfDNA) from 1-6ml cell-free bodyfluids (such as plasma, serum). The purified DNA is suitable for direct use indownstream applications such as PCR, real-time PCR, Biochip analysis and NGS.
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