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Invitrogen™ Human Cot-1 DNA™
Description
Includes
This DNA is supplied at 1mg/mL in 10mM Tris-HCl (pH 7.4), 1mM EDTA.
Human Cot-1 DNA is commonly used to block nonspecific hybridization in microarray screening. It can also be used to suppress repetitive DNA sequences for the direct mapping of human DNA or mapping genomic clones to panels of somatic-cell hybrids for chromosome localization by Southern blotting.
About Cot-1 DNA
Human Cot-1 DNA is placental DNA that is predominantly 50 to 300 bp in size and enriched for repetitive DNA sequences such as the Alu and Kpn family members. The amount supplied per package is sufficient for 5–10 Southern or 500 in situ hybridizations.
Performance and quality testing
Purity and DNA size are verified by agarose gel electrophoresis. Concentration is verified spectrophotometrically by diluting Human Cot-1 DNA 1:100 in 50mM NaOH and using the conversion factor 0.033μg/μL/A260. Other methods used to determine concentration may yield varying results.
Array CGH, Cell Analysis, Cellular Imaging, ChIP-on-Chip, Chromatin Biology, Fluorescence In Situ Hybridization, Gene Expression Analysis and Genotyping, Genotyping and Genomic Profiling, In Situ Hybridization (ISH), Microarray Analysis, RNAi, Epigenetics and Non-Coding RNA Research
Order Info
Shipping Condition: Approved for shipment on Wet or Dry Ice
Specifications
Specifications
| Product Type | DNA QC |
| Content And Storage | This DNA is supplied at 1 mg/ml in 10 mM Tris-HCl (pH 7.4), 1 mM EDTA. Store at -20°C. |
| Reagent Type | Cot-1 DNA |
| Shipping Condition | Approved for shipment on Wet or Dry Ice |
| Species | Human |
| Product Line | Cot-1 DNA |
| For Use With (Application) | Block Non-Specific Hybridization |
| Quantity | 500 μg |
Frequently Asked Questions (FAQs)
For Southern blot hybridizations, add 50 µg of COT-1 DNA (at 10 µg/µL) to 50 µL of 20X SSC, 25 µL distilled water and 20 µL of a solution containing 0.1 M NaCl, 0.1 M Tris-HCl (pH 7.4). 0.01 M EDTA, and 1% SDS to the probe for each 25 to 500 ng of probe. For in situ hybridizations, combine genomic probe with the proper amount of COT-1 DNA such that the final concentration of COT-1 DNA is 0.3 µg/µL for cosmid, plasmid, and lambda probes; or, at 1 µg/µL for Alu PCR probes. Ethanol precipitate and resuspend in a half-volume of 100% formamide. Add a half-volume of 20% dextran sulfate in 2X SSC (prewarmed to 75 degrees C) and mix well. Denature mix by heating to 75 degrees C for 5 min. Incubate at 37 degrees C for 5 to 15 min.
Probes can be labeled with 32P by random primer or nick translation procedures using the Random Primers DNA Labeling System (Cat. No.18187-013) or Nick Translation System (Cat. No. 18160-010). Biotinylated COT-1 DNA can be prepared by nick translation with the BioNick Labeling System (Cat. No. 18247-015) or by the BioPrime DNA Labeling System (Cat. No. 18094-011). Improved results can be obtained when the COT-1 DNA is first ligated to itself to provide an optimum template.
Human Cot-DNA-Fluorometric QC has been quantitated by fluorometry; this provides more accurate concentration measurements for higher molecular weight products. Mouse Cot-1 DNA blocking reagent is recommended for experiments when blocking of repetitive mouse DNA is required.
Cot-1 DNA blocking reagent blocks repetitive sequences such as SINEs (short interspersed elements), LINEs (long interspersed elements), and sequence homology among members of the same gene family when added to the hybridization solution.
Better results are obtained when Cot-1 DNA blocking reagent is used due to reduction in cross hybridization, which leads to cleaner, more sensitive aCGH microarray experiments and FISH assays.
For Research Use Only. Not for use in diagnostic procedures.
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