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The primer sequences used for PCR amplification were as follows (restriction sites indicated in lower case): HK2 3'UTR BglII FW: 5'-GGGagatctGGAGGGATGAGAGTGGCTTA-3' HK2 3'UTR XhoI RV: 5'-GGGctcgagAATGACAACATCTTCACTAGACTGAG-3' The miR-143 8mer seed site, TCATCTCA, in the 3'UTR of HK2 was converted into TCAT GACA using the QuikChange site-directed mutagenesis kit (Stratagene, La Jolla, CA, USA).
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His move follows restrictions imposed on union activity in Whitehall by the Cabinet Office minister, Francis Maude, on Monday.
The EC said its move, which follows restrictions introduced unilaterally by the UK in November 2010, was designed to forward the European Union's stated mission to abolish the death penalty around the world.
The next is an example that does not follow restriction (3), since (s)(a) is not just a type variable: instanceShow ((s)(a)) (Rightarrow )Show (Sized(s)(a))... Instances that do not follow these restrictions are common in Haskell programs, specially in the presence of multi-parameter type classes.
Following restriction endonuclease digestion of intact Tetrahymena Mt DNA, the terminal restriction fragments were slightly "smeared" due to the variable length of the telomeres.
Following restriction endonuclease digestion, DNA samples were electrophoresed through 0.7% agarose gels, and transferred to Nytran SuPerCharge membrane (Schleicher and Schuell) according to standard procedures [72].
Following restriction digestion with XhoI and PacI, these APP, tau, SOD1 and TXN 5' UTR sequences were inserted between the XhoI and PacI sites of php15 to generate pGL4.15cmvAPP5, pGL4.15cmvtau5, pGL4.15cmvSOD1.5 and pGL4.15cmvTXN5 respectively.
Following restriction and adaptor ligation, there followed two successive rounds of PCR amplification.
Following restriction enzyme digestion the DNA was treated with sodium bisulfite without denaturing the double-stranded fragments.
Enzymes were then inactivated by heating to 75°C for 15 min. Following restriction and adaptor ligation there followed two successive rounds of PCR amplification.
Following restriction digest with Bsa I and mung bean nuclease treatment, the linearized plasmid was used in a run‐off in vitro transcription reaction.
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