Clontech PT3139-1 User Manual - Page 17
Browse online or download pdf User Manual for Laboratory Equipment Clontech PT3139-1. Clontech PT3139-1 20 pages. Advantage-hf pcr kit
Print PageV. Troubleshooting Guide continued
D. Dealing with contamination
Contamination most often results in extra bands or smearing. It is important
to include an H
template) in every PCR experiment to determine if the PCR reagents,
pipettors or PCR reaction tubes are contaminated with previously amplified
targets.
If possible, set up the PCR reaction and perform the post-PCR analysis in
separate laboratory areas with separate sets of pipettors.
Laboratory benches and pipettor shafts can be decontaminated by
depurination. Wipe surfaces with 1N HCl followed by 1N NaOH. Then
neutralize with a neutral buffer (e.g., Tris or PBS) and rinse with H
It is advisable to use one of the commercially available aerosol-free pipette
tips.
There is an enzymatic method for destroying PCR product carryover
(Longo et al. , 1990). It involves incorporation of dUTP into the PCR
products and subsequent hydrolysis with uracil-N-glycosylase (UNG).
When performing PCR directly on phage plaques or bacterial colonies,
failure to isolate single plaques or colonies will also produce multiple bands.
TEL:415-424-8222 or 800-662-CLON
FAX:415-424-1064 or 800-424-1350
O control (i.e., a control using H
2
Technical Support
CLONTECH Laboratories, Inc.
O instead of the DNA
2
Protocol # PT3139-1
Version # PR76834
O.
2
page
17