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BLAST (EMBL, Flybase) and domain searches (SMART) were performed to identify Drosophila proteins that contain the mucin-associated SEA, vWD4, EGF and cystein-knot domains, and identified proteins were manually scanned for the presence of PTS repeats.
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A steam generator tube rupture (SGTR) accident analysis for SMART was performed using the TASS/SMR-S code.
Their songs were snappy, their lyrics were smart, and they were performed by genuine outsiders too (photos of a knock-kneed Cocker playing celebrity five-a-side in Select magazine proved that).
In order to gain more material, 30 ng of normalised cDNA were used for 100 μl PCR and 7 cycles of PCR amplification with SMART PCR primer were performed as suggested by Evrogen (http://www.evrogen.com/kit-user-manuals/Trimmer-2.pdf).
SMART and Pfam analysis were performed to remove those putative pseudogenes and incorrect annotated genes, and then resulted in 91 members recognized by either SMART (Sm00356) or Pfam (PF00642).
For PCR reactions, PCR and the real-time amplification monitoring for the PNA-LNA PCR clamp were performed using Smart Cycler II (Cepheid Sunnyvale, CA, USA).
The single-crystal X-ray diffraction measurements were performed using Bruker SMART APEX II CCD diffractometer.
Real-time PCR (RT-PCR) analyses were performed with a Smart Cycler real-time PCR instrument (Cepheid, Sunnyvale, CA).
The siRNA-mediated interference experiments for eEF1γ expression were performed by transfecting SMART pool-specific or non-specific control pool double-stranded RNA oligonucleotides (Dharmacon) using Lipofectamine 2000.
First strand cDNA synthesis and amplification were performed using the SMART RACE cDNA Amplification kit (Clontech Laboratories Inc., CA, USA) following the manufacturer's instructions.
To obtain the UTR sequences, both 3' and 5' RACE were performed with the SMART RACE kit (TaKaRa Co, Japan) following the manufacturer's instructions.
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