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Genomic DNA was used as a control template in parallel PCR reactions to confirm the expected electrophoretic pattern of the retroposed sequences.
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PCR reactions were performed with experimental and control templates in parallel.
The challenge is to detect the flashes of light from each unique DNA template while sequencing numerous templates in parallel.
PCR reactions were performed with experimental and control templates in parallel and PCR products were run on 7% polyacrylamide gels and quantified using the Kodak Gel Logic 2000 imaging system (Kodak, Rochester, NY).
PCR products were quantitated with Sybr green (Invitrogen Corporation) and compared with dilutions of cloned plasmid template amplified in parallel.
The PCR products were quantitated by hybridization with a Taqman probe (genomic coordinates 4218 4189) and converted to genome copies by comparison with a standard curve of cloned plasmid template amplified in parallel.
qPCRs were performed in triplicate for each of the 23 cDNA pools along with a no template control in parallel for each gene.
Now, similar to the process that we modeled in this work in silico, we can envision evolution of such a system to shorten ribozyme-template interactions in parallel with evolution of the decoding apparatus (culminating in the modern ribosome) compensating for the loss of stability and discriminative power for such shortened interactions.
Moreover, as the placement template generator runs in parallel with the layout-aware loop, it has no impact on the overall execution time.
A negative control without template was run in parallel to assess the overall specificity of the reaction.
Enhancing the cationicity of the feleucin template may result in parallel enhancement of potency and spectrum of action.
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