Cells and Tissue DNA Isolation Kits
![Cells and Tissue DNA Isolation Kit](/sites/default/files/styles/norproduct_imagekit/public/images/product_images/53100-Cells-and-Tissue-DNA-Isolation-Kit-WEB.jpg?itok=zch50fij)
For research use only and NOT intended for in vitro diagnostics.
Aperçu du produit
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.
Détails
Supporting Data
Figure 1. Isolation of High Quality Genomic DNA from Tissue. Genomic DNA was isolated from 15 mg of liver tissue using Norgen's Cells and Tissue DNA Isolation Kit. Triplicate samples were used (lanes 1-3) and yielded genomic DNA of the highest quality and integrity. Lane M is the Norgen UltraRanger 1kb DNA Ladder. For all purified DNA, 15 µL of each 200 µL elution were resolved on a 1X TAE, 1% agarose DNA gel.
Figure 2. Isolation of High Quality Genomic DNA from 0.1 up to 3 million mammalian cells. Genomic DNA was isolated from 0.1 x 106 to up to 3 million HeLa cells using Norgen's Cells and Tissue DNA Isolation Kit. Triplicate samples were used from each cell input and yielded genomic DNA of the highest quality and integrity with linear increase in yield with increasing cell input. Lane M is the Norgen UltraRanger 1kb DNA Ladder. For all purified DNA, 15 µL of each 200 µL elution were resolved on a 1X TAE, 1% agarose DNA gel.
Figure 3. Yield of Genomic DNA from 0.1 x 106 up to 3 x 106 mammalian cells. Genomic DNA was isolated from 0.1 up to 3 million HeLa cells using Norgen's Cells and Tissue DNA Isolation Kit. Triplicate samples were used from each cell input and 25 µL of the 200 µL elution were diluted in 475 µL of nuclease-free water, and DNA concentrations were measured using the UltraSpec 2100 Pro (Fisher Scientific). Linear increase in DNA yield was obtained when increasing the cell input from 0.1 x 106 - 3 x 106 HeLa cells.
Figure 4. Purified DNA Can be Amplified in a Real-time PCR (TaqMan) Reaction. Genomic DNA was isolated from 0.1 x 106 to 3 x 106 of HeLa cells using Norgen's Cells and Tissue DNA Isolation Kit. Five µL of the DNA from each 200 µL elution was used in a real-time PCR reaction (total reaction volume of 20 µL) with GAPDH TaqMan probe and primers. The real-time PCR was successful in amplifying the GAPDH gene from all the isolated DNA from the different cell inputs with a decreasing Ct value with increasing the cell input. This indicates that the kit provides high quality DNA in each case and is free of PCR inhibitors making it well suited to sensitive downstream applications.
Figure 5. Resolution of DNA isolated from Animal Tissue and Mammalian Cells. DNA was isolated from 20 mg animal tissue (beef) and 1 x 106 HeLa cells using Norgen's Cells and Tissue DNA Isolation Kit (Magnetic Bead System), and Norgen's Cells and Tissue DNA Kit (column format, Cat. 53100) as a control. The DNA isolated with the Magnetic Bead System (B) was then compared with the column method (C). For evaluation, 10 µL from each 200 µL of elution were run on 1X TAE 1.2% agarose gel. As it can be seen, Norgen's Magnetic Bead System (B) was able to isolate high yields of DNA from both the tissue and cells. M = Norgen's HighRanger DNA Ladder.
Figure 6. Comparison of A260/280 and A260/230 ratios. DNA was isolated from 20 mg animal tissue (beef) and 1 x 106 HeLa cells using Norgen's Cells and Tissue DNA Isolation Kit (Magnetic Bead System), and Norgen's Cells and Tissue DNA Kit (column format, Cat. 53100). The DNA isolated with the Magnetic Bead System was then compared with the column method for A260/280 and A260/230 rations. DNA isolated using Norgen's Cells and Tissue DNA Isolation Kit (Magnetic Bead System) showed a comparable DNA quality to the column method (Cat. 53100).
Figure 7. High Quality DNA was confirmed by Real-time PCR. DNA was isolated from 20 mg animal tissue (beef) and 1 x 106 HeLa cells using Norgen's Cells and Tissue DNA Isolation Kit (Magnetic Bead System). The high quality of the purified DNA was confirmed by Real-time PCR using 2 µL (circle), 4 µL (triangle) and 8 µL cross) of animal tissue DNA or HeLa DNA (total PCR reaction volume was 20 µL) to detect 5s rDNA or GAPDH from animal tissue and HeLa cell respectably. No PCR inhibition was observed from the all different PCR template amounts, indicating the excellent quality of animal tissue and HeLa DNA for downstream applications.
Figure 8. DNA was Isolated from 1 x 106 HeLa Cells Using Norgen's Cells and Tissue DNA Isolation 96-Well Kit (Magnetic Bead System). For evaluation, 10 µL from each 100 µL of elution were run on 1X TAE 1.2% agarose gel. As it can be seen, Norgen's Cells and Tissue DNA Isolation 96-Well Kit (Magnetic Bead System) was able to isolate consistent and high yields of DNA from HeLa cells. M = Norgen's HighRanger DNA Ladder (Cat. 11900).
Figure 9. High Quality DNA was Confirmed by Real-Time PCR. DNA was isolated from 1 x 106 HeLa cells using Norgen's Cells and Tissue DNA Isolation 96-Well Kit (Magnetic Bead System). The high quality of the purified DNA was confirmed by Real-time PCR using 2 µL of HeLa DNA (total PCR reaction volume was 20 µL) to detect GAPDH from HeLa cell. No PCR inhibition was observed indicating the excellent quality of HeLa DNA for downstream applications.
Kit Specifications
|
|
Maximum Input |
20 mg of animal tissue |
Column Binding Capacity | > 50 μg |
Average Yield | 8 μg (from 1 x 106 HeLa Cells) 10 μg (from 10 mg kidney) |
Elution Volume | 50 - 200 μL |
Analyte Purified | Genomic DNA, mitochondrial DNA, viral DNA |
Format | Spin Column |
Time to Complete 10 Purifications | 30 min (cells) and 90 min (tissue) |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment. The kit contains a ready-to-use Proteinase K, which is dissolved in a specially prepared storage buffer. The buffered Proteinase K is stable for up to 1 year after the date of shipment when stored at room temperature.
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 |
Documentation
(57300) Cells and Tissue DNA Isolation Micro Kit - Protocol (50 preps)
(59100) Cells and Tissue DNA Isolation Kit (Magnetic Bead System) - Protocol (50 prep)
(62500) Cells and Tissue DNA Isolation 96-Well Kit (High Throughput Magnetic Bead System) - Protocol (2 x 96-well)
Comparing Different DNA and RNA Quantification Methods for Biological Samples with Low Nucleic Acid Abundance
Determination of the DNA Molecular Weight (MW) from different Norgen Columns and Isolation Methods