RAPD Problems - Control lanes with bands
szcooley at chip.ucdavis.edu
Thu Jul 13 09:37:56 EST 1995
ANTHONY WILSON (002233w at axe.acadiau.ca) wrote:
: I've been performing RAPD reactions using primers from the UBC lab
: and have been running a control with the same stock as my tests (Primer,
: taq, buffer, dNTPs, H20) replacing the DNA with sterilized water, yet I am
: consistently getting banding patterns in these lanes WITHOUT DNA... I have
: replaced the H20, dNTPs, taq and buffer and yet the bands are as strong as
: ever! Is this a common setup for a control lane?
: These banding patterns differ from reaction to reaction and from primer
: to primer, yet bands appear in control lanes even when some of the test
: lanes are clean... Has anyone had any similar experiences and/or any
: suggestions as to where in the protocol this error might be introduced??
: Any help would be greatly appreciated.
: Tony Wilson
The problem is that DNA is contaminating your TAQ. It is in all the TAQ
polymerase sources at some level and is generally not a problem. The
level is generally low enough that when template DNA is added the "real"
bands are preferentially amplified. Later, Mike
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