Housekeeping

... simultaneously. I have read somewhere that one way for normalizing PCR data is to divide the Ct of the gene of interest by the Ct obtained from a housekeeping/reference gene (Ct inter. / Ct refer.). However, I am not sure how to interpret the resulting values. Is it true that the larger the ratio, ...

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... is that I did PCR with RNA and cDNA simultaneously and did gel electrophroesis. I obtained very interesting results which corresponding bands of housekeeping gene were detected when RNA was used as template but not the target gene. We thought of possibly of genomic DNA contamination but when ...

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... gene products: for some proteases, very bright bands. for some transcription factors highly expressed at the same time, no bands at all. for a housekeeping gene in the species (ubiquitin), no bands. the regions of these gene i am amplifying are all 2kb, cycler conditions and PCR rxn mix followed ...

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... came up with a set of primers. Now i have to calculate the approximate length of the amplified sequence. with the following criteria to be noted : housekeeping gene has to be amplified at the same time, same sample etc, target mRNA should produce a DNA distingushable with housekepper and taq polymerase ...

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... providing a picture as to which spliced variants occur. High throughput normalisation (subtractive hybridisation would eliminate 90% of the known housekeeping as well (or DNA chip screening, but you do miss the advantage of the splice variants). 2. development stage limitation: some of the genes ...

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