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myprayer: 金币+1, 赠人玫瑰，手有余香，分子生物期待您更多精彩。 2016-01-13 18:13:33
Your results have been identified the key problem, which was the DNA sample containing high level of polysaccharides that binding or co-precipitate with DNA. The evidences were 1) positive amplification at low concentration which could reduce the contaminates, 2) highlight strength above the agarose-gel well which was the high level of polysaccharides that binding or co-precipitate with DNA.
Therefore, you can purify your DNA samples using high salt TE protocol below.
1. Add 400 ul of high salt TE (1M NaCl in TE buffer) +2 ul of RNaseA (10mg/mL)
2. Incubate at 37 degrees Celsiu for one hour, mixing from time to time until the pellet dissolves
3. add 2 volume of cold 100% ethanol, put at -20 degree Celsiu for 20 minutes
4. Spin at max speed for 15 minutes
5. Wash pellet with 1ml of cold 75% ethanol
6. Spin at max speed for 5 minutes
7. Repeat wash
8. Dry pellet and resuspend in 100 ul of TE (or more, as you prefer)
Or you can re-extract your DNA samples with 3% CTAB DNA extraction protocol, which separates high quality DNA in our labs.
![16S扩不出来，到底是模板问题还是引物问题？]()
3% CTAB DNA Extraction Protocol.jpg
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