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One possibility for the similarities between the antisense and control groups can be the temporal expression of Tbx2. Perhaps the time when antisense blocking of Tbx2 would have the greatest affect on hair follicle outgrowth is earlier than 13.5 dpc. If this is the case, then the plan for hair follicle outgrowth is determined, and manipulation would not cause any noticeable effects. The genes that require the TBX2 transcription factor would already be regulated, and thus manipulation of Tbx2 after this point would not change the functioning of that gene. Another possibility lies in the treatment of the whisker pads with antisense. It is possible that the experiment was not successful in blocking the Tbx2mRNA. This possibility can be reaffirmed or eliminated by performing further diagnostic agarose gels. After treating the whisker pads and allowing them to culture for a short time, their RNA would be isolated and converted to cDNA. This would then be multiplied by PCR and run in a gel. There should be a weakening, or lack of the Tbx2bands in the antisense cultures, and a maintaining of this band in the controls. Running this type of diagnostic to ensure the blocking of mRNA is the next logical step in the research plan. If the incubations with 200nM antisense are shown to be ineffective in blocking the function of Tbx2,higher concentrations of antisense might be attempted as well as attempting to use different solutions to introduce the antisense oligonucleotides.

Although no conclusions regarding the role Tbx2plays in hair follicle development can be made according to the results garnered by this one experiment, the experiment was successful in providing a sound basis from which other experiments can be designed.

©Cebra-Thomas, 2000

Last Modified: 05 May 2000

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