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Natural Science Forum / Biology / Biology / October 2006hi
Hi, I am trying to do PCR and after doing electrophoresis and I notice that there is contamination in negative control. I repeated the same many times, by changing the samples under sterile conditions. I just want to know is there any possibility to know the reason for contamination. The samples I am currently working on are related to transgenic mice. regards, sai. >Hi, >I am trying to do PCR and after doing electrophoresis and I notice >that there is contamination in negative control. Which negative control? >I repeated the same >many times, by changing the samples under sterile conditions. I just >want to know is there any possibility to know the reason for >contamination. Have you recently done a similar procedure in the same lab facilities without seeing the contamination? Are others in the same lab doing PCR without contamination? Are you doing the PCR work in a clean room in which no other DNA work is allowed? bob >The samples I am currently working on are related to transgenic mice. > >regards, >sai. Hi, I did PCR in the same lab many times and there was no contamination at all. But this time I don't know I am repeating the expt by taking a great care..I don't understand where the fault lies. In fact, I changed the completed solutions too. Regards, Sai. > >Hi, > >I am trying to do PCR and after doing electrophoresis and I notice [quoted text clipped - 18 lines] > >regards, > >sai. >Hi, > >I did PCR in the same lab many times and there was no contamination at >all. But this time I don't know I am repeating the expt by taking a >great care..I don't understand where the fault lies. In fact, I changed >the completed solutions too. It can be hard to sort out. It is good that you have been successful before; at least that gives you something of an existence theorem. On the other hand, perhaps you were lucky, and really need more stringent procedures. PCR labs often take quite extreme precautions, since DNA contamination is so easy. You might check some PCR books or web sites and read as much as you can about contamination. Maybe take this as an opportunity to improve basic procedures. (I have no idea what your system is.) You have done some of the right trouble-shooting -- but obviously not enough. One thing you might do is to see if you can identify what the contaminating DNA is. Even simpler... are you getting the same contaminant band in independent replicates of the same expt? Again, looking for clues. bob >Regards, >Sai. [quoted text clipped - 21 lines] >> >regards, >> >sai. > my friends: I can tell you why pcr needing a clean suroundings. As it's contamination can cause large disarster for you research,even the little contimanate of hair and tips. any thing beyound the sample may product fault result. |
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