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Invitrogen™ PureLink™ Genomic DNA Mini Kit
Description
The PureLink™ Genomic DNA Mini Kit enables high-yield, high-purity, genomic DNA (gDNA) extractions from a wide variety of sample types. Use this single kit for genomic DNA purification from blood, tissues, cells, bacteria, swabs, and blood spots, with a familiar silica-based, microcentrifuge spin-column format. Features of the PureLink™ Genomic DNA Mini Kit include:
• Kit flexibility—one kit works with a variety of sample types and sizes
• Ultraclean gDNA—minimal contamination of DNA product means successful downstream application
• Improved design—optimized spin-column design and buffer formulation for better yield and purity
Multiple sample sources, one kit
The PureLink™ Genomic DNA Mini Kit can be used with many different sample types, each with its own optimized protocol outlined in the manual.
Optimized components and protocols
The PureLink™ Genomic DNA Mini Kit includes parts and protocols that improve upon the easy and familiar spin-column methods currently in use. Columns are newly configured to bind and release DNA more efficiently. Each sample source uses a specialized lysis procedure and an optimized lysis buffer formulated to enhance proteinase K activity and eliminate protein contamination. A chaotropic salt buffer promotes stable binding of DNA to the column resin, and powerful wash buffers remove all traces of protein and salt. DNA is eluted in a low-salt buffer to allow for pH stabilization of the DNA in storage.
Higher yields and purity
With PureLink™ Genomic DNA Kits you can expect high yields of high-purity gDNA (determined by A260/A280 measurements), no matter the sample type, from bacteria to tissues to blood to cultured cells. The enhanced PureLink™ Lysis and Wash Buffers help to consistently deliver purified gDNA samples with absorbance reading ratios of ∼1.9 for both A260/A280 and A260/A230, indicating no detectable contamination from carryover of protein or guanidine.
Order Info
Shipping Condition: Room Temperature
Specifications
Specifications
| Content And Storage | • 2.0 mL PureLink™ Genomic Lysis/Binding Buffer • 1.8 mL PureLink™ Genomic Digestion Buffer • 2.0 mL PureLink™ Genomic Wash Buffer 1 • 1.5 mL PureLink™ Genomic Wash Buffer 2 • 2.0 mL PureLink™ Genomic Elution Buffer • 0.4 mL RNase A (20 mg/mL) • 0.2 mL Proteinase K (20 mg/mL) • 10 PureLink™ Spin Columns with Collection Tubes • 20 PureLink™ Collecion Tubes (2.0 mL) Store all components at room temperature. |
| Isolation Technology | Silica Spin Column |
| Scale | Mini |
| Sample Type | Bacteria, Blood, Cells, Tissue |
| For Use With (Application) | Genotyping, Real-Time Quantitative PCR (qPCR), Southern Blotting, Sequencing, PCR |
| High-throughput Compatibility | Not High-throughput Compatible (Manual) |
| Prep Scale | 100 μL to 1 mL Blood Samples (Medium-Scale) |
| Product Line | PureLink |
| Product Type | Genomic DNA Mini Kit |
| Quantity | 10 Preps |
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Frequently Asked Questions (FAQs)
We would recommend using our PureLink gDNA Mini Kit or our ChargeSwitch gDNA Mini Bacteria Kit (Cat. No. CS11301), which can isolate both gram-positive and gram-negative bacteria. No centrifugation or filtration steps are necessary using this simple one-tube protocol. Read more about bacterial DNA extraction here (http://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/dna-extraction/genomic-dna-extraction/bacteria-dna-extraction.html).
We offer TRIzol reagent that will allow isolation of DNA and RNA from the same sample. Alternatively, we have the following method that has been validated by our R&D team; for sequential isolation of gDNA and total RNA from the same sample. This method involves using 2 of our kits: 1) PureLink RNA Mini Kit (Cat. No. 12183018A, 12183020, 12183025) and 2) PureLink Genomic DNA Mini Kit (Cat. No. K182002, K182000, K182001).
The protocol is detailed below:
Before starting:
- Label all spin columns and buffers from each kit with kit names to prevent confusion.
- Prepare lysis buffer and wash buffers according to the protocol from each kit.
1. Preparing lysates:
- Add 300 µL of lysis buffer (from Purelink RNA Mini Kit, beta-mercaptoethanol added) to cell or tissue sample, lyse the cells as recommended.
2. DNA isolation:
- Load all of the lysate directly onto a Purelink gDNA column (from PureLink Genomic DNA Mini Kit), save flow-through for RNA isolation.
- Centrifuge the Purelink gDNA column at 10,000 x g for 1 min.
- Wash the Purelink gDNA column with 500 µL of Wash Buffer 1 (from PureLink Genomic DNA Mini Kit, ethanol added), centrifuge at 10,000 x g for 1 min.
- Add 500 µL of Wash Buffer 2 (from PureLink Genomic DNA Mini Kit, ethanol added), centrifuge at maximum speed for 3 min to dry the membrane.
- Add 100 µL of Elution Buffer (from PureLink Genomic DNA Mini Kit), incubate at room temperature for 1 min and centrifuge at 10,000 x g for 1 min (yield can be increased if an optional second elution step is added).
- This is purified gDNA.
3. RNA isolation:
- To the above saved flow-through, add same volume of 70% ethanol, mix well and load the lysate/ethanol mix (including all precipitates) onto an RNA spin cartridge (from Purelink RNA Mini Kit).
- Centrifuge at 12,000 x g for 15 sec. Discard flow-through.
- Wash the RNA spin cartridge with 700 µL of Wash Buffer 1 (from Purelink RNA Mini Kit, ethanol added), centrifuge at 12,000 x g for 15 sec.
- Wash twice with 500 µL of Wash Buffer 2 (from Purelink RNA Mini Kit, ethanol added). After the second wash, centrifuge at 12,000 x g for 1 min to dry the membrane.
- Add 50 µL of RNase-free water onto the RNA spin cartridge, incubate at room temperature for 1 min and centrifuge at 12,000 x g for 2 min (yield can be increased if an optional second elution step is added).
- This is purified RNA.
We recommend the following DNA isolation kits:
- RecoverAll Total Nucleic Acid Isolation Kit for FFPE (Cat. No. AM1975)
- Ion Ampliseq Direct FFPE DNA Kit (Cat. Nos. A31133, A31136)
- MagMAX FFPE DNA/RNA Ultra Kit (Cat. No. A31881)
- PureLink Genomic DNA Mini Kit (Cat. Nos, K182000, K182001, K182002)
For Research Use Only. Not for use in diagnostic procedures.