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Although there are many reconstruction algorithms for localization microscopy, their use is hampered by the difficulty to adjust a possibly large number of parameters correctly. We propose SimpleSTORM, an algorithm that determines appropriate parameter settings directly from the data in an initial self-calibration phase. The algorithm is based on a(More)
We report an improved variant of mKeima, a monomeric long Stokes shift red fluorescent protein, hmKeima8.5. The increased intracellular brightness and large Stokes shift (∼180 nm) make it an excellent partner with teal fluorescent protein (mTFP1) for multiphoton, multicolor applications. Excitation of this pair by a single multiphoton excitation wavelength(More)
Here, we report on a novel PCR targeting-based strategy called 'PCR duplication' that enables targeted duplications of genomic regions in the yeast genome using a simple PCR-based approach. To demonstrate its application we first duplicated the promoter of the FAR1 gene in yeast and simultaneously inserted a GFP downstream of it. This created a reporter for(More)
1 Purpose The purpose of this RFC is to introduce a standardized framework for the engineering of customizable non-ribosomal peptide synthetases (NRPS) and their application for in vivo and in vitro synthesis of short non-ribosomal peptides (NRPs) of user-defined sequence and structure. 2 Relation to other BBF RFCs BBF RFC100 updates RFC57 describing the "(More)
We have developed OnTarget, a pipeline which is designed to evaluate next generation sequencing data after sub-genomic enrichment. It is able to process single-end and paired-end sequence reads as well as single-end bisulfite data. Sequence reads can be sorted by barcodes and are mapped against the genome. Only reads which map once are processed,(More)
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