...interaction partners (PIPs), the FP fragments form the complete FP because the PIPs they’re tethered to ...endogenous PIPs is expressed at increasing levels alongside your BiFC constructs. If your PIPs actually ... YFP) is reunited by the interaction between two PIPs, resulting in a fluorescent readout.   Applications...aberrant subcellular localization of the studied PIPs. The straightforward nature of BiFC also makes it...protein construction Fusing FP fragments to your PIPs requires careful selection of FP fragments and, ...some information about the structures of the tested PIPs. Many split FPs have been validated for BiFC (Kodama...Kodama and Hu, 2012). Having a rough idea of how your PIPs might interact will allow you to determine where...

...TEF2 Yeast Piper 60931 pPL5608_TEF1*-Cre_TRP1 Cre expressed at low levels mutant TEF1 Yeast Piper 60932 pPL5606... Yeast Piper 60933 pPL5628_TEF1*-Cre_MET15 Cre expressed at low levels mutant TEF3 Yeast Piper 61391 pCS2...

...facilitates screening by allowing a multichannel pipette to be used for the remainder of the steps in step...well) to a 96-well PCR plate using a multichannel pipette. Centrifuge PCR plate at 400 x g for 5 min and ...clone present. Resuspend cells in 200 μl of media. Pipette mix to detach cells. Plate 100 μl each into two...

...CRISPResso is a robust and easy-to-use computational pipeline that enables accurate quantification and visualization...

...TCAACATTGGATATCTATagagctttacagctagTTTATCCCAGTGTCCAGA IPIP27A TAL3296 & TAL3297 TGAACGAGAGGAGTGTGGcacactatgccacatgCAACTCTCCTCCAGACAA...

... monitor the entire pipeline. Benefits of Nextflow for bioinformatics pipelining In many use-cases, including...optimize our pipeline. After reviewing and testing several platforms for engineering pipelines, we selected...management system As novices to creating a robust data pipeline that would be reliable and available for our teams... could orchestrate the steps into a continuous pipeline. We also recognized the software system that we...ours at Addgene, the overall purpose of a data pipeline is to perform several sequential computational...usage of the scripts needs to be customized in the pipeline for the specific needs of the user. Thus, a key..., a workflow management system for scientific pipelines, including bioinformatics. We loved the parallels...

...the larval gut of bagpipe mutants vaguely resemble a bagpipe and historically, bagpipes were made using ... and bagpipe, two genes important for mesoderm differentiation Now, let's turn tinman and bagpipe, two... Cookie Monster, Spaghetti Squash, Tinman, and Bagpipe. How does a gene get its name? Often times, a gene...becomes muscles and connective tissues. tinman and bagpipe are both homeobox genes that are part of a genetic... the dorsal mesoderm, is necessary to activate bagpipe which is localized to segmental clusters of the... to have a heart (Following the Tinman, 2016). Bagpipe on the other hand, is important in the formation...into the muscles of the gut. Embryos that lack bagpipe have severely disrupted visceral mesoderm and subsequently...

...200 ul pipettor is set to 200 ul, wiped down with 70% ethanol, and a box of sterile truncated pipet tips...the pipettor, placed into wells, and partially dislodged from the agarose plug by rapid pipetting, as ...rapidly punch out cell clones with sterile truncated pipet tips. Indeed, an experienced operator can pick 48.... We highly recommend the use of an electronic pipettor with variable spacing (such as the Viaflo from...

...curve, avoid pipetting small volumes (< 2 µL) or volumes too large for a standard micropipette (> 1,000 µL...2. When preparing a dilution series, use a new pipette tip for each step and mix the samples well by vortexing...readout. When preparing a standard curve, change pipette tips and mix thoroughly by inversion or vortexing...series, use consistent dilution factors, change pipette tips regularly, and mix samples very well.  Use...

...into a medium-throughput pipeline for epitope tagging. Using our 96-format pipeline, we can routinely achieve...to contact me at [email protected]. Keep pipetting and happy genome editing! Pooran Dewari is a ... them to study brain tumour biology. When not pipetting, he can be seen daydreaming about sunny days in...Kalantzaki M et al. 2018. An efficient and scalable pipeline for epitope tagging in mammalian stem cells using...

... into your data. Try different types of pipettes and pipetting methods to find what’s most consistent ...plate in a way that will tell you if you have pipetting errors on the edges of the plate. Compare the ...multiple configurations can help you identify pipetting or evaporation errors. And if time allows, you...BioRender.   Passive Reference Dyes Whatever your pipetting method, some variation will occur. A passive reference...

...source workflow orchestration software for data pipeline processing (Nextflow), cloud infrastructure, and...collection of scripts evolved into a single, organized pipeline and accelerated the processing of our sequencing... has used Nextflow to improve and redesign our pipeline. In it, they break down the unique challenges ...

...scientists washed and reused materials such as glass pipettes or inoculation loops. While single-use is now ...a single use. So how about washing and reusing pipette tips in a high-throughput and efficient way? Grenova... devices that are compatible with a variety of pipette tips. This is part of a cultural change in how ...

...screens together and expedited the drug discovery pipeline. Basics of drug discovery Present drug discovery...potential targets by CRISPR can accelerate the drug pipeline. High-throughput CRISPR screens can be used to... aids in multiple stages of the drug discovery pipeline.   CRISPR and drugs in the future While ...

...as a device fits on the deck, and it’s easy to pipette samples in and out of it, you can set up a procedure...using electronic multichannel and single channel pipettes. These can speed up any project in which you need... to do a lot of aliquoting. Simply program the pipette with how much total sample you need and the volume...

...of a reference sequence) using a bioinformatics pipeline developed at Addgene. The assembled plasmid sequence...request. Otherwise, they continue through the QC pipeline to resolve any issues.  Catching and addressing...arise due to errors in the sequencing and assembly pipeline. There are regions of plasmids that are particularly...

...Have a dedicated bench with a dedicated set of pipettes for ddPCR set-up. Prepare the master mix in a ...the bench and all consumables with 10% bleach.  Pipette slowly If you are using a manual droplet generator...automated droplet generator, you should avoid pipetting too quickly when making your virus dilutions. ...

... to poor lab techniques. These include reusing pipettes for multiple cell lines rather than using disposable...aerosols that are generated (while talking or pipetting) during working in the laminar airflow are also... are tightly packed with seals. Use disposable pipettes to avoid cross-contamination. Make sure you keep...

... appreciated the creative uses of old posters, pipette tip boxes, and more (stay tuned for a few blog ...sustainability in the lab!). The @ladiesinlabcoats used a pipette tip box and microcentrifuge tubes to make Eppi ...