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Aliquots of 1 µl PCR products amplified with four different colour dye labelled primers and three different product sizes were pooled together to reduce cost and increase efficiency.
Multiplex microsatellite amplification was carried out using QIAGEN Type-it™ microsatellite PCR Kit with fluorescent dye labelled forward primers [ 46].
Sequencing reactions using universal infra red fluoresecent dye labelled M13 sequence oligonucleotides were then carried out on the PCR products.
Forward and reverse primers were end dye labelled with Dyomics fluorescent tags DY-681 (700 nm absorption) and DY-781 (800 nm absorption), respectively.
Therefore, instead of synthesizing one specific fluorescently labelled primer for each SSR marker, only a dye labelled M13 primer is needed.
Fluorescent dye labelled M13 forward primer and a marker specific reverse primer were used to generate fluorescent-labelled PCR product as previously described [ 72].
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All TaqMan Real Time PCR reactions were performed using using FAM-labelled PINK1 probe and VIC/TAMRA-dye labelled Endogenous control Human RPLPO or GAPDH (Applied Biosystems).
Hybridisations were performed using 15 20 μg of Cy-dye labelled amplified-RNA (aRNA) from each infected cell line.
Total RNA was Cy-dye labelled and hybridized on miRNA microarray chips as previously described [ 24].
Mutations were identified in individual lines from the durum and bread wheat libraries by evaluating infrared-dye labelled PCR products digested at mismatched sites in heteroduplexed DNA [ 30].
This microarray was hybridized with cyanine-dye labelled cRNA probes synthesized from RNA from female or male adults of H. contortus.
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