Laboratory Centrifuges

4 days ago

Let's chat Feedstocks, Cellular Chassis, Fermentation Systems, DSP, and Packaging. Who's trying to manufacturing commodity-competitive products with microbes in vats? I'm open to sharing my experience and expertise in designing & building consumer hardware, electronic devices, chemical processing equipment, liquid handlin...

21 days ago

This is a quick run-through of the protocol to make dilution spot plates for S. cerevisiae. This assay is great for identifying broad fitness defects but isn't quantitative. Note that temperatures for temperature sensitive strains will need adjustment from the standard 30°C. Even if you aren't aware of any temperature-rela...

about 1 month ago

This protocol served me well through my PhD and postdoc years. Dilute the final product at least 1/10 for use in PCR. Spin down 50-100 uL packed volume of yeast cells Resuspend in 200 uL TE + 1% SDS and move the cells to a 1.5ml screw-top tube if possible Add 200 uL of glass beads (eyeball this: just pour in beads until th...

about 2 months ago

The concept of proteolytic targeting chimera (PROTAC) was first proposed by Crews and Deshaies laboratory in 2001, the team successfully achieved the degradation of methionyl aminopeptidase 2 (MetAp-2) by PROTAC method. Since then, PROTACs targeting androgen receptor (AR) and estrogen receptor (ER) have appeared one after a...

2 months ago

I just stumbled upon an extremely cheap and simple way to clean up PCR products without the need for columns or fancy reagents. Here's my slight tweak of this wonderful Open Wetware entry. PCR Precipitation Buffer To a 15mL tube, add 3g PEG 3350. (I use Miralax brand which works well) Add 2.19g NaCl. Fill to 12mL with dist...

3 months ago

This is my go-to alakline lysis miniprep protocol that does not require a kit or silica columns. It's based on the tried and true protocols of yesteryear and brought to my attention a few years ago by J. Leon (@jesussaur) on Twitter. I've been using it ever since and have successfully sent plasmids out for sequencing via pl...

10 months ago

I used this method to make electrocompetent NEB turbo E. coli cells. This protocol is based on the one from Barrick labs which is linked below. https://barricklab.org/twiki/bin/view/Lab/ProtocolsElectrocompetentCells One colony of NEB turbo E. coli was taken from a petri dish and inoculated into 5 mL of LB broth, in a 1...

almost 2 years ago

Gel extractions can be a low yield affair, but overnight incubation over a spin column can increase yields. This protocol details a modified QIAquick Gel Extraction protocol. QIAquick Gel Extraction Kit Cut the desired band out of an agarose gel using a scalpel. Weigh the slice in a snap cap tube and add 3 volumes of QG ...

almost 2 years ago

This protocol details a typical setup for screening for transformed cells in the process of cloning a gene following a typical cloning procedure. An alkaline phosphotase is use to ensure there is no self ligation of the vector. α-Complementation - Many shuttle vectors contain an ampicillin resistance gene so only successfu...

Laboratory Centrifuges

Building a Biomanufacturing Knowledge-base

Building a Biomanufacturing Knowledge-base

Dilution spot assay for yeast

Dilution spot assay for yeast

"Quick and dirty" S. cerevisiae gDNA prep protocol

"Quick and dirty" S. cerevisiae gDNA prep protocol

A Review of Proteolytic Targeting Chimeras

A Review of Proteolytic Targeting Chimeras

Ultra-cheap PCR Precipitation using PEG and Salt

Ultra-cheap PCR Precipitation using PEG and Salt

Columnless Old-School Miniprep

Columnless Old-School Miniprep

Electrocompetent NEB turbo E. coli prep

Electrocompetent NEB turbo E. coli prep

Higher Yield Gel Extractions

Higher Yield Gel Extractions

X-Gal Blue White Screening

X-Gal Blue White Screening