pcr prob

[ChenHA]

Aawara Chowdhury wrote:
> In <1203946852.17846.0 from proxy01.news.clara.net>,
>  ChenHA <hzhen from freeuk.com> wrote:
> 
>> Aawara Chowdhury wrote:
>>
>>> P.S. You don't have to be so rude, just because someone points out
>>> an incongruence in your "observation".
>>>
>> Geez, now you are truly stupid.  Why put the quotation marks on 
>> observation?  Are you suggesting that observations were wrong, or false, 
>> or I have mistaken a reflection of a light bulb to be a band?  What's 
>> incongruent about my observation?  Observations are observations, 
>> science comes out of observation, idiot.  I have shown that you made 
>> unwarranted assumption on primer concentration and length, and you still 
>> insist on piling on casting doubt on my "observation".  Stupid people 
>> deserve all the insult they get.
> 
> Your "observation" is incongruent with everything that has been published
> about the function of ethidium bromide.  You "observe" that ethidium
> fluoresces "brightly" when bound to single-stranded oligonucleotides.
> That either means that your oligonucleotides were not single-stranded
> (odd for PCR primers), or that there's an error with your "observation".
> And yes, that is precisely why I find your "observation" perplexing.
> And unfortunately for you, science is not just about making an "observation";
> it also requires being prepared to subject an observation to review.  
> 
> And my, don't we like calling people stupid.  Makes you feel intelligent,
> does it?

Certainly from the evidence more intelligent than you.  Which bit of the 
"unwarranted assumption on primer concentration and length" don't you 
understand?  What is the length of primers used?  You don't know (mine 
are usually more than 30 but less than 60, very occasionally 80 or 
more).  Do they fold back?  Possibly, or perhaps they form loops, or 
whatever (and very probably, these have no relevance to what happened in 
hot reactions compare to what might happen in a room temperature gel). 
Do they form primer dimer in the reaction, maybe, maybe not, but from my 
own experience they usually aren't primer dimers (and they may indeed 
form dimer in gel, but that may have no relevance to what happened in 
the reaction).  You are arguing from no experience, read a few things 
then spouting off about things you have no experience of.
Idiot.






> 
> AC