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Questions related from Rachel Andrea Chea
I was extracting genomic DNA from Streptomyces the previous day and left my gDNA in the oven (55 degrees Celsius) to dry at the very last step. I had to rush off for a seminar and completely...
09 September 2019 7,792 2 View
I remember my mentor saying before that she decided to refrigerate her gentamycin stock solution instead of freezing it since that was the recommended storage condition. Now, I'm dealing with...
08 August 2019 9,643 5 View
I intend to clone in some genes into my vector (pIJ10257) using Gibson Assembly but I'm not sure if I should add a terminator at the end of the genes before cloning them into the vector. Is there...
07 July 2019 2,030 1 View
To cure my Streptomyces strain of a plasmid, I used 5-FC as a negative selection marker. So, cells without the plasmid (which are the ones I'm looking for) will be able to grow on agar plates...
07 July 2019 5,441 2 View
I am currently using the CRISPR base editing system in bacteria. Unfortunately, I'm not getting any clones with edited bases (not even off-target) but i'm not sure why. I'm quite sure the plasmid...
05 May 2019 1,828 4 View
I've extracted my gDNA from Streptomyces today, on a Friday but it's pretty late and I can't stay late today. Is it okay for me to keep my extracted gDNA over the weekends at -20 degrees Celsius...
04 April 2019 5,318 10 View
I've just done ligation for my plasmid and my next step would be to perform heatshock transformation on Top10 E. coli cells. Some of my colleagues recommended me to use 2 uL to transform my cells...
01 January 2019 1,568 4 View
I've been trying to clone a ~100 bp insert into a ~12000 bp vector backbone. Basically, what I've done is to do the standard things (restriction digestion, PCR cleanup/gel extraction, ligation,...
12 December 2018 9,413 13 View
I have PCR-ed some DNA samples which I need to perform gel extraction on. These DNA samples will subsequently be used for restriction digestion and ligation over the next week. I was advised to...
12 December 2018 3,991 7 View
Currently, my project involves the preparation of Pseudomonas aeruginosa mutants and one of the steps involves triparental mating to transform P. aeruginosa with the mutant plasmid. My question...
03 March 2018 7,975 4 View
