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A major benefit of this novel design is that it allows reverse shaft rotation.
The availability of C. albicans genome sequences allows reverse genetics strategies to be employed on a large scale.
RNA content of lentiviral vectors was quantified with a one-step RT-qPCR that allows reverse transcription (RT) and amplification to take place in the same reaction tube.
Total RNA of all donors was pooled, and for each microarray experiment, 150 μg was reverse-transcribed and amplified by SMART™-PCR (Clontech, Mountain View, CA, USA), a technology that allows reverse transcription (RT) of small amounts of total RNA and subsequent amplification of the entire cDNA.
Sensing of HIV-2 in DCs is attributable to Vpx, which allows reverse transcription to proceed efficiently, and a suboptimal CA structure, which permits the cytosolic DNA sensor cGAS to gain access to reverse transcribed cDNA and to trigger a host response.
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A tagged-mutagenesis approach allowed reverse engineering of improved strains, revealing two genes, mutB and cobA, in the primary metabolic branch for methylmalonyl-CoA utilization.
By allowing reverse transcription to proceed, we find that DCs detect HIV-1 in distinct phases, before and after virus integration.
An additional side-effect is that the approach exceeds the requirements of the GNU Lesser General Public License by allowing a software vendor to easily provide a limited amount of source code, rather than potentially releasing source code for large portions of their product suite, or allowing reverse engineering of potentially large portions of their proprietary product.
The other is by mutations that increase phosphorolysis leading to removal of already incorporated chain-terminating inhibitors from the DNA, thereby allowing reverse transcription to continue.
Cell line RNA was used in combination with the stem-loop Multiplex primer pools (Applied Biosystems), allowing reverse transcription of 48 different miRNA in each of 8 RT pools.
Total RNAs were then isolated using the High Pure RNA Isolation Kit Protocol (Roche) and subjected to RT-PCR to generate cDNA using p dT 15 primer (Roche) and Expand Reverse Transcriptase (Roche), allowing reverse transcription of total sample mRNA.
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