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Questions related from Omar Ocegueda
Hello everyone, I had some plasmids synthesized by TWIST. I transformed them into homemade chemically competent NEB-stable. During the outgrowth, I accidentally grew them at 37C instead of 30C,...
15 January 2025 4,184 1 View
I'm fairly new to making homemade comp cells and have done so with some mild success. What is generally an acceptable way of resuspending comp cells during wash steps? I've been told to swirl the...
01 August 2024 8,746 0 View
I'm currently trying to capture a biosynthetic gene cluster using Transformation-Associated Recombination (TAR) in yeast. After identifying positive yeast clones, I extract, via an alkaline lysis...
19 April 2024 8,619 4 View
I was making competent cells the other day with a culture volume of 100 mL (diluted from an O/N) in a 500 mL flask. Starting at 1 h, I started measuring OD to check when the cultures reached...
03 April 2024 2,908 3 View
I noticed that a lot of plasmid annotations tend to find ColE1 origins, but I can never usually find the b. subtilis origin of replications from annotations.
22 March 2024 7,293 2 View
I'm trying to build a cosmid for B. subtilis, but there are no commercially available cosmids with a gram-positive ori already installed. I was thinking of obtaining a commercial cosmid (SuperCos...
20 April 2023 4,614 0 View
Hello All, I am performing a Golden Gate assembly, and I've noticed that I've been seeing pale blue colonies alongside really dark blue colonies. My system is designed so that when the golden...
01 January 1970 1,895 3 View
