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Six weeks after intrasplenic injection, the mice were sacrificed and autopsy performed to check for formation of liver metastases.
There have been very few measurements of rate constants reported for GSK-3, as the vast majority of studies use simple qualitative endpoint assays, for example western blotting to check for formation of phosphorylated substrates, to measure activity.
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It has been postulated that if sense and antisense RNAs are present in the same cell, they can form dsRNA, but it is difficult directly check for the formation of dsRNA in vivo.
A negative control (no-template control) was included in each primer assay to check for the formation of primer-dimers.
After 1 week, media were observed under a microscope to check for axon formation and enlargement of nuclei (all data not shown).
Samples were selected at random following T and M reactions and subjected to agarose gel electrophoresis to confirm PCR specificity and to check for primer dimer formation.
After each run, a melt-curve was produced in 1°C increments starting at 60°C to check for primer dimer formation.
Restriction digests and gel electrophoresis of DNA minipreps (Qiagen) were used to check for the correct formation of PCR products and Entry and Destination clones.
Embryos were raised to adulthood and checked for tumor formation.
Sequences were checked for chimera formation with the CHECK_CHIMERA software of the Ribosomal Database Project [101].
Asynchronous blood-stages of P. berghei NK65 (WT) and Plrx were maintained in NMRI mice and checked for gametocyte formation and exflagellation of microgametes prior to mosquito feeding.
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