PEG3-bis(Amino-Tri-(Propargyl-PEG8-ethoxymethyl)-methane) is large molecule with two sets of three terminal alkynes joined together by a short PEG linker. The alkynes are most frequently used in copper click chemistry with azides to form stable triazoles with the target compound. The PEG linkers as well as numerous amide bonds provide high aqueous solubility to this compound, which may alter DMPK of this compound.
PEG3-bis(Amino-Tri-(Propargyl-PEG8-ethoxymethyl)-methane) is large molecule with two sets of three terminal alkynes joined together by a short PEG linker. The alkynes are most frequently used in copper click chemistry with azides to form stable triazoles with the target compound. The PEG linkers as well as numerous amide bonds provide high aqueous solubility to this compound, which may alter DMPK of this compound.
This kit provides a rapid, single column method for the isolation and purification of total RNA (including miRNA) and proteins sequentially from a single sample of cultured animal cells, tissues, blood, bacteria, yeast, fungi or plants. The total RNA and proteins are both column purified in under 25 minutes using a single column.
Purified RNA is of a high quality and yield, and is suitable for NGS, RT-qPCR and microarrays.
This kit eliminates DNA efficiently using a gDNA removal column.
Proteins are eluted in buffer and are ready for downstream applications such as Western Blots, Mass Spec and ELISA. The proteins will not require precipitation, resuspending of pellets, or any further cleaning.
This kit is ideal for researchers who are interested in studying the transcriptome and proteome of a single sample, such as for studies of microRNA profiling, gene expression including gene silencing experiments or mRNA knockdowns, studies involving biomarker discovery, and for characterization of cultured cell lines. Norgen’s RNA/Protein Purification Plus Kit is especially useful for researchers who are isolating macromolecules from precious, difficult to obtain or small samples such as biopsy materials or single foci from cell cultures, as it eliminates the need to fractionate the sample. Furthermore, analysis will be more reliable since the RNA and proteins are derived from the same sample, thereby eliminating inconsistent results. The purified macromolecules are of the highest purity and can be used in a number of different downstream applications.
Protocol
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| Kit Specifications | |
| Maximum Column Binding Capacity | 50 μg for RNA |
| Maximum Column Binding Capacity | 200 μg for Protein |
| Maximum Column Loading Volume | 650 μL |
| Size of RNA Purified | All sizes, including small RNA (< 200 nt) |
| Time to Complete 10 Purifications | 30 minutes |
| Average Yields*: HEK 293 Cells (1 x 106 cells) HEK 293 Cells (1 x 106 cells) Liver (15 mg) Liver (15 mg) | 10-15 μg RNA 70-100 μg protein 30-35 μg RNA 100-150 μg protein |
* Average yields will vary depending upon a number of factors including species, growth conditions used and developmental stage.
Storage Conditions and Product Stability
The Protein Loading Dye should be stored at -20°C after the addition of DL-Dithiothreitol (DTT). All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.
| Component | Cat. 48200 (50 preps) |
|---|---|
| Buffer SKP | 40 mL |
| Wash Solution A | 38 mL |
| Elution Solution A | 6 mL |
| Wash Solution C | 30 mL |
| Binding Buffer A | 8 mL |
| Elution Buffer C | 4 mL |
| Protein Neutralizer | 4 mL |
| Protein Loading Dye | 2 mL |
| gDNA Removal Columns | 50 |
| RNA/Protein Purification Columns | 50 |
| Collection Tubes | 150 |
| Elution Tubes (1.7 mL) | 100 |
| Product Insert | 1 |
Norgen’s Cells and Tissue DNA Isolation Kits are designed for the rapid preparation of genomic DNA from cultured cells as well as various tissue samples and urine. The purified genomic DNA is fully digestible with all restriction enzymes tested, and is completely compatible with PCR and Southern Blot analysis.
Cells and Tissue DNA Isolation Kit (Spin Column)
Purification is based on spin column chromatography as the separation matrix. Norgen’s columns bind DNA under optimized salt concentrations and release the bound DNA under low salt and slightly alkali conditions. The protocol can be completed in approximately 30 minutes for cells and within 90 minutes for tissues. Each kit contains sufficient materials for 50 preparations.
Cells and Tissue DNA Isolation Micro Kit (Micro)
Optimized for small inputs of cells and tissues, such as Laser-Captured Microdissection (LCM). Purification is based on spin column chromatography as the separation matrix. Norgen’s columns bind DNA under optimized salt concentrations and release the bound DNA under low salt and slightly alkali conditions. Preparation time for a single sample is approximately 60 minutes, and each kit contains sufficient materials for 50 preparations.
Cells and Tissue DNA Isolation Kit (Magnetic Bead System)
Purification is based on the use of magnetic beads that bind DNA under optimized binding conditions. Norgen’s Cells and Tissue DNA Isolation Kit (Magnetic Bead System) allows for the isolation of genomic DNA from various types of animal tissues or cell samples. Preparation for 10 purifications is approximately 40 minutes of hands-on time.
Cells and Tissue DNA Isolation 96-Well Kit (High Throughput Magnetic Bead System)
Purification is based on the use of magnetic beads that bind DNA under optimized binding conditions. Norgen’s Cells and Tissue DNA Isolation 96-Well Kit (Magnetic Bead System) allows for the isolation of genomic DNA from various types of animal tissues or cell samples. The Cells and Tissue DNA Isolation 96-Well Kit (Magnetic Bead System) also can be integrated with a robotic automation system.
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| Kit Specifications | |
| Maximum Cells and Tissue Input | 20 mg of animal tissue 3 x 106 cells |
| Average Yield* | 8-10 µg (20 mg of animal tissue) 8-12 µg (3 x 106 cells) |
| Average Purity (OD260/280) | 1.8 – 1.9 |
| Time to Complete 10 Purifications | 40 minutes hands-on time (Cat. 59100) 50 minutes hands on time (Cat. 62500) |
* Average DNA yield will vary depending on the donor
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature (15 – 25°C). This kit is stable for 1 year after the date of shipment.
| Component | Cat. 53100 (50 preps) | Cat. 57300 (50 preps) | Cat. 59100 (50 preps) | Cat. 62500 (192 preps) |
|---|---|---|---|---|
| Lysis Buffer B | 20 mL | 20 mL | 20 mL | 1 x 40 mL 1 x 20 mL |
| Solution WN | 18 mL | 18 mL | 18 mL | 55 mL |
| Wash Solution A | 18 mL | 18 mL | – | – |
| Elution Buffer B | 30 mL | 8 mL | 15 mL | 1 x 30 mL 1 x 15 mL |
| Proteinase K | 1.2 mL | 1.2 mL | 1.2 mL | – |
| Proteinase K in Sotrage Buffer | – | – | – | 4 mL |
| Magnetic Bead Suspension | – | – | 2 x 1.1 mL | 8.5 mL |
| 96-Well Plate | – | – | – | 2 |
| Spin Columns | 50 | – | – | – |
| Micro Spin Columns | – | 50 | – | – |
| Collection Tubes | 50 | 50 | – | – |
| Elution Tubes (1.7 mL) | 50 | 50 | 50 | – |
| 96-Well Elution Plate | – | – | – | 2 |
| Adhesive Tape | – | – | – | 2 |
| Product Insert | 1 | 1 | 1 | 1 |
The series of DNA Size Selection Kits (Magnetic Beads) were developed for DNA size selection using magnetic beads. A total of 11 kits are available, with different selection ranges spanning from 50 bp to over 10 kb. The kits provide a simple and quick approach for the enrichment of a specific range of DNA fragments. The kit workflow allows double-sided or single-sided size selection for specific size cutoffs.
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DNA size selection is a selective capture of DNA fragments of a specific range of size for next-generation sequencing (NGS) library preparations, PCR, ChIP assay, DNA ligations, endonuclease digestions, adapter removal, and other genomics and molecular biology applications. DNA size selection is preferred after NGS library prep in most of the cases. The NGS library preparation is related to the quality of the sequencing data. Precise NGS library size selection can increase sequencing efficiency, improve data quality, and reduce costs.
There are two types of sequencing technologies: short-read sequencing and long-read sequencing. Short-read sequencing uses DNA libraries that contain small insert DNA fragments of similar sizes, usually several hundred base pairs. The sequencing efficiency can be improved if the DNA size selection is in the right range. Cat.# 20104S and 20104L are the best kits for NGS library size selection of illumina paired-end 100 (PE100) sequencing with 100-200 bp library inserts; Cat.# 20105S and 20105L are the best kits for NGS library size selection of illumina paired-end 150 (PE150) sequencing with 150-300 bp library inserts; and Cat.# 20106S and 20106L are the best kits for NGS library size selection of illumina paired-end 300 (PE300) sequencing with 300-600 bp library inserts.
Long-read sequencing uses a large DNA fragment as input and makes very long reads. Usually, library size selection is preferred to remove smaller fragments. Cat.# 20110S and 20110L are the best kits for long-read sequencing size selection with DNA sizes >5 kb, and Cat.# 20111S and 20111L are the best kits for long-read sequencing size selection with DNA sizes >10 kb.
The magnetic beads technology uses paramagnetic particles, also known as SPRI (Solid Phase Reversible Immobilization) beads, to bind DNA reversibly and selectively. DNA fragments can be size-selected and purified by changing the properties of the magnetic beads or SPRI beads. The magnetic beads can easily separate the beads-binding DNA from the contaminants and unwanted components in the samples. The samples after DNA size selection are free of contaminants such as buffer components, enzymes, proteins, salts, dNTPs, primers, and adapters. Our proprietary magnetic beads reagents improve yield, selectivity, and reproducibility.
Specific DNA fragments at a certain length range can be purified simply using magnetic separation with different beads components, avoiding tedious and time-consuming gel extraction and column-based purification. The magnetic beads method is popular for common DNA size selection, including library size selection. The first beads-binding step, referred to as the right-side clean-up, removes large DNA fragments. The large DNA fragments are bound to the beads and are discarded. The desired DNA fragments in the supernatant are transferred to a new well, and new beads are added to the supernatant for the second beads-binding, referred to as the left-side clean-up. The double-size selected DNA fragments are eluted after ethanol rinsing.
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A single clean-up is needed for DNA size selection with large fragments. In this case, only the large DNA fragments are bound to the beads. The selected larger DNA fragments are eluted after ethanol rinsing.
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