PEG13-bis(Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane) is large molecule with two sets of three terminal alkynes joined together by a large PEG linker. The alkynes are most frequently used in copper click chemistry with azides. The PEG linkers as well as numerous amide bonds provide high aqueous solubility to this compound, which may alter ADME of this compound.
PEG13-bis(Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane) is large molecule with two sets of three terminal alkynes joined together by a large PEG linker. The alkynes are most frequently used in copper click chemistry with azides. The PEG linkers as well as numerous amide bonds provide high aqueous solubility to this compound, which may alter ADME of this compound.
Sample type purification kit guide
The 16S V3-V4 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V3-V4 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 V3-V4 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 V3-V4 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.
| 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 V3-V4 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. 70400 (24 preps) | Cat. 70410, 70420, 70430, 70440 (96 preps) |
|---|---|---|---|
| Amplicon PCR (PCR 1) | MGX Master Mix | 330 µL | 1,320 µL |
| 16S V3-V4 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 |
This product is suitable for rapid extraction of 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.
Specifications
| Features | Specifications |
| Main Functions | Isolation total DNA from blood, buffy coat, tissue and other samples |
| Applications | Second generation sequencing, PCR, real time PCR, etc. |
| Purification method | Polydisperse magnetic beads |
| Purification technology | Magnetic beads technology |
| Process method | Manual or automatic |
| Sample type | Anticoagulant blood, concentrated blood, buffy coat, lymphocytes and cultured cells |
| Sample amount | Whole blood :< 200μl; Saliva / swab:< 400μl; Tissue :< 20mg |
| Yield | 0.1 – 50μg |
| Elution volume | |
| Time per run |
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 | IVD3102 |
| Purification Times | 200 |
| MagPure Particles | 5 ml |
| Proteinase K | 100 mg |
| Protease Dissolve Buffer | 10 ml |
| Rnase A | 40 mg |
| Buffer ATL | 60 ml |
| Buffer AL | 60 ml |
| Buffer BD* | 20 ml |
| Buffer BW1* | 110 ml |
| Elution Buffer | 30 ml |
Storage and Stability
Proteinase K, RNase A, MagPure Particles should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 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
This product is suitable for rapid extraction of 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.
