Omar Ocegueda

5 Questions 2 Answers 0 Followers

Questions related from Omar Ocegueda

What is the best practice for resuspending cell pellets during competent cell preparation (both electro and comp)?

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 8,141 0 View

Troubleshooting Transforming and isolating a large plasmid in E. Coli?

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 8,561 4 View

How to accurately measure the OD of larger bacterial cultures?

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 2,852 3 View

How to check if a plasmid can be replicated in E coli and/or B. subtilis?

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 7,159 2 View

Are cosmids easy to genetically modify?

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 4,562 0 View