Over the last month our DNA sequencing has hit some strange snags.
It’s difficult to troubleshoot a process when you’ve been doing it the same way for so long and previously has just worked fine!
This time I cooked up some NZy broth to grow cultures in before mini-prepping them. The main difference between the broth we normally use and the NZY seems to be using NA amine and adding MgCl2 and MgSO4.
I also found a nice mac app to use to align the sequences in order to verify the DNA was definitely my gene. Check out Serial Cloner. It has lots of useful features.
Tomorrow I should be able to mutate! Wish me luck!
NZY+ Broth (per Liter)
10 g of NZ amine
5 g of yeast extract
5 g of NaCl
Add deionized H2O to a final volume of 1 liter. Adjust to pH 7.5 using NaOH.
Add the following filer-sterilized supplements prior to use:
12.5 ml of 1 M MgCl2
12.5 ml of 1 M MgSO4
20 ml of 20% (w/v) glucose (or 10 ml of 2 M glucose)