Patrick Astarita
4 days ago
Question

Building a Biomanufacturing Knowledge-base

Building a Biomanufacturing Knowledge-base

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...
_._darachm_._ {LastName}
8 days ago
MethodPro Lab Tip

Warnings, solutions, and mysteries regarding extraction/PCR from large amounts of yeast stationary-phase genomes

Warnings, solutions, and mysteries regarding extraction/PCR from large amounts of yeast stationary-phase genomes

This post is about some perplexing and difficult to get rid of contaminant that accumulates in yeast, especially stationary phase samples, that inhibits PCRs in a hard to predict way. Now I'm no biochemist, so I don't have a solution for you, but I do have a dirty hack that should let you do large batches of extracts that a...
Sebastián Sosa Carrillo
17 days ago
Method

Quick and easy bioengineering procedures: Yeast (S. cerevisiae) colony PCR

Quick and easy bioengineering procedures: Yeast (S. cerevisiae) colony PCR

Hi all, This is the last post in a series of 4 in which I want to share a quick and easy to use procedure for engineering a yeast strain. The idea is to go from the cloning stage, right after doing a PCR (or order) to get the DNA you want to clone, up to the process of transforming the yeast and selecting the positive clon...
Karissa Milbury
21 days ago
Method

Dilution spot assay for yeast

Dilution spot assay for yeast

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...
Karissa Milbury
about 1 month ago
Method

"Quick and dirty" S. cerevisiae gDNA prep protocol

"Quick and dirty" S. cerevisiae gDNA prep protocol

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...
Sebastián Sosa Carrillo
about 2 months ago
Method

Quick and easy bioengineering procedures: Yeast (S. cerevisiae) transformation

Sam
about 1 year ago
Method

Materials and Methods

Materials and Methods

Strains, media and storage The Saccharomyces cerevisiae strains used in this work belong to the CEN.PK family (Entian and Kötter 2007; Salazar et al.2017) and are listed in Table 1. The strains were grown at 30°C in 500 mL flasks containing 100 mL chemically defined medium (synthetic medium, SM) (Verduyn et al.1992) or yea...
Sam
about 1 year ago
Method

Materials and Methods

Materials and Methods

Strains, growth conditions and storage The S. cerevisiae strains used in this study (Table ​(Table1)1) share the CEN.PK genetic background (Entian and Kötter 2007; Nijkamp et al. 2012). Shake flask cultures were grown at 30 °C in 500 mL flasks containing 100 mL synthetic medium (SM) (Verduyn et al. 1992) with 20 g·L−1 gluc...
Sam
about 1 year ago
Method

Materials and Methods

Amos Fung
almost 2 years ago
Method

TOPO TA Cloning