This is a method for low-cost electrotransformation of electrocompetent NEB turbo E coli cells using a homemade electroporation apparatus. The original paper upon which this method is based can be found here: https://doi.org/10.1371/journal.pbio.3000589 My method for preparing electrocompetent E coli can be found here: h...
Hi folks, I found a nice overview over the strength of various BioBricks promoters https://jbioleng.biomedcentral.com/articles/10.1186/1754-1611-3-4 I'm especially interested in finding a constitutive promoter (family) so I can test a range of promoters with different known strengths, for example the BioBricks J23100 prom...
I am working with carrots, which are, in general, very easy to culture. Just pop them on MS media with 2,4-D and they will produce callus within a month. My only problem is that my carrots seem to have a LOT of endophytic bacteria that take over very quickly. What can I do to suppress or kill these bacteria? I plan on using...
We know there's tons of troubleshooting information on Twitter so we've spoken with our community council and decided to start up a campaign to start archiving all sorts of experimental resolutions from across #sciencetwitter to add to our "lab-rary" using the tag #scifound. I've provided guidelines below to have an idea of...
Hello all, I am performing PCR on about 1000 bacterial isolates (lysis solution only, no DNA extraction) for a culturomics project. PCR will amplify V1-V3 of 16S gene and then sending these out for sequencing to identify each isolate. I am searching for alternatives to running a small aliquot of each PCR product on a large...
Hi everyone, I've received a custom oligo pool from Twist Bioscience and they provided 70ng of it. We talked to one of the reps and they said not to resuspend it to any less than 10ng/μL, which means I'll be adding less than 7μL of buffer, which is obviously not a lot. I was wondering if anyone had any practical tips for h...
Genetic EngineeringGeneticsMicrobiologyMolecular Biology Oligo Pool Twist Bioscience Twist Oligo Pools
First of all, @Michael Paulsmeyer had a great post a while back on how to get as much DNA from a standard miniprep as possible (https://www.scifind.net/methods/efficient-miniprep-spin-column-k). The variance in my personal results from minipreps is large, though. To make sure I always get a high concentration of DNA, I use ...
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Find your community. Ask questions. Science is better when we troubleshoot together.
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