_._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...
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...
Amos Fung
almost 2 years ago
Method

Cas9 Condition Optimization

Cas9 Condition Optimization

This protocol runs several in vitro Cas9 cleavage conditions for identifying the optimal setup for a specific target (in this case the Sub1 gene). Cas9 Cleavage Cas9 nuclease reactions can be set up as described in the table below. For identifying cleavage conditions, use a combination of 1X and 2X Cas9 reactions along wi...
Guy Rohkin
almost 2 years ago
Method

Extracting DNA from Silkworm is Difficult potentially due to endogenous DNAses