Alator - Model M205 -Cot I, Human DNA

Applications:

• In situ suppression (CISS) hybridizations
• Hybridization to micro-arrays
• Other In situ hybridizations
• Filter hybridizations

Size: Bulk
Concentration: >= 1.1 mg/ml
Storage Buffer: 10 mM Tris-HCl (pH 7.4), and 1 mM EDTA.

Quality Control:

• Average fragments size: 50-300 bp;
• A260/A280 ration: 1.6 – 2.0
• Amount of genomic (non-repetitive DNA): less than 2%.
• COT I DNA and the raw material is tested for the absence of HIV1,2 RNA, HCV RNA, HBV DNA

Storage and Handling: Store at -20°C, before use the frozen product should be at 4°C slowly thawed

For Southern blot hybridizations: add 50 μg of COT-1 human DNA (@ 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 human DNA such that the final concentration of COT-1 human 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 halfvolume 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 °C for 5 min. Incubate at 37 °C for at least 5 and up to 15 min.