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PCR band smear

PCR band smear - Protocols and Methods Forum

PCR band smear - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 11-13-2006, 01:43 AM
muhammad yasir
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Default PCR band smear



i am running PCR unsing PCR product as templet and facing the problem of smear at extactly the same position aroung of desired band size. Any sugestion for this problem
with thanks
Yasir


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  #2  
Old 11-14-2006, 11:37 AM
Domo
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Default PCR band smear

Ta ==> 60C for 30 sec

increase annealing temperature

"muhammad yasir" <[Only registered users see links. ]> wrote in message
news:[Only registered users see links. ].ne t...


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  #3  
Old 11-14-2006, 12:47 PM
Duncan Clark
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Default PCR band smear

Historians believe that in newspost
<[Only registered users see links. ].net> on Sun, 12 Nov 2006,
muhammad yasir <[Only registered users see links. ]> penned the following literary
masterpiece:

How much are you diluting the original PCR product and how many cycles
are you then doing ?

Duncan
--
I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.
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  #4  
Old 11-14-2006, 07:19 PM
Damien
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Default PCR band smear

You can reduce smearing by using less DNA polymerase, decreasing the
number of cycles, or decreasing extension time. Increasing template
concentration might help as well.

You may also try another DNA polymerase. I use Rainbow
([Only registered users see links. ]).

Damien



muhammad yasir wrote:
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  #5  
Old 11-15-2006, 12:41 PM
Duncan Clark
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Default PCR band smear

Historians believe that in newspost <ejd4sn$ng3$[Only registered users see links. ].edu> on Tue,
14 Nov 2006, Damien <[Only registered users see links. ].edu> penned the following
literary masterpiece:

Ah, self advertising.

Duncan
--
I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.
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band , pcr , smear


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