DNase treatment, cDNA synthesis, primer design and SYBR Green I RT-PCR were carried out as described . In brief, 2 μg of each total RNA sample was treated with the RQ1 RNase-free DNase according to the manufacturer's instructions (Promega). Treated RNA samples were desalted (to prevent carry over of magnesium) before cDNA synthesis using Microcon-100 spin columns (Millipore). First-strand cDNA was synthesized using random hexamers and SuperscriptII reverse transcriptase according to the manufacturer's instructions (Invitrogen), and subsequently diluted with nuclease-free water (Sigma) to 12.5 ng/μl cDNA. RT-PCR amplification mixtures (25 μl) contained 25 ng template cDNA, 2x SYBR Green I Master Mix buffer (12.5 μl) (Applied Biosystems) and 300 nM forward and reverse primer. Reactions were run on an ABI PRISM 5700 Sequence Detector (Applied Biosystems). The cycling conditions comprised 10 min polymerase activation at 95°C and 40 cycles at 95°C for 15 sec and 60°C for 60 sec. Each assay included (in duplicate): a standard curve of four serial dilution points of SK-N-SH or IMR-32 cDNA (ranging from 50 ng to 50 pg), a no-template control, and 25 ng of each test cDNA. All PCR efficiencies were above 95%. Sequence Detection Software (version 1.3) (Applied Biosystems) results were exported as tab-delimited text files and imported into Microsoft Excel for further analysis. The median coefficient of variation (based on calculated quantities) of duplicated samples was 6%.
Suitable for beginners and experienced music editors, this free program comes with a vast sound library containing effects and royalty-free music. It also supports the use of plugins and saves audio in popular audio formats. Users who wish to access more tools have the option of upgrading to the premium version of WavePad download. Having said that, the free program includes almost all features that you might find in other similar programs like Audacity, Lexis Audio Editor, and Free Audio Editor. It is also Windows 11 compatible.
Developed by NCH Software, WavePad lets users manipulate audio tracks, add effects, and change sample rates. The feature-rich application supports a range of file formats, including MP3, VOX, WMA, and FLAC, among others. It also comes with a sound-effect library that includes royalty-free music and sound effects.
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This free sound editor includes many audio editing effects such as amplify, normalize, equalizer, envelope, reverb, and more. Easily edit audio files with cut, copy, paste, delete, insert and several others. Audio restoration features includes noise reduction and click pop removal. 2b1af7f3a8