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Although the region-specific CGG PCR failed to amplify the sample with >60 repeats (a female premutation, Fig. 1a), the TP PCR accurately sized this sample (CGG = 31, 69 and 91, Fig. 1b).
Magnetic streptavidin beads are used to bind to the biotinylated probes, the nontargeted portion of the genome is washed away, and the polymerase chain reaction (PCR) is used to amplify the sample, enriching the sample for DNA from the target region.
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To ensure they had a large enough sample of the virus to get results, they amplified the sample through a series of DNA sequences which trigger replication.
If one or both fragments were not amplified, the sample was considered to bear fully integrated virus, otherwise the sample was designated as containing mixed forms of the virus.
The other primer combinations either non-specifically amplified the samples or did not amplify a product at all.
Using the sPMCA technique as described (1 ), we amplified the samples in 3 8 tubes, and we used Western blot to analyze the proteinase K treated sPMCA products (2 ).
We amplified the samples ("original," "complex," and "free") using NEBNext Fast DNA Library Prep Set reagents with 1 nM DNA PCR reaction mix, 10 μM each of IDT Ion Torrent A primer, and truncated P1 primer (for PCR conditions, see Supplementary Materials Note 2).
Because the cRNA yield from the 1 μg pooled sample test was low, we decided to amplify the smaller samples.
They amplified the tiny sample into a large quantity of DNA using a standard technique called whole genome amplification.
All the test primer pairs were verified by amplifying the control sample and sequencing the PCR products.
PCR reactions were normalized by amplifying the same sample of cDNA with primers specific for glyceraldehyde-3-phosphate dehydrogenase, GAPDH.
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