[DIYbio] Four questions about reliability of a wet lab action

                        

Greeting reader(s):

I posted this question buried in a deeper strand, I think maybe it has to bubble up to get noticed enough to snag opinions, answers; maybe some direct experience 'stories':

Question began 'sort of' to to Sean Sullivan, possibly but really a general question

Here are four questions about creating a gene edit and expression, ex Yeast, even e coli, etc... Some plate and Petri dish wetlab style work. All are about framing expectations and resources, the assumption is the technology is 'best we can get', and/or almost isn't the point, anyway.

Reference Situation:

*) Gene BP count is 3K NT, promoter, tails, introns if any etc known to work generally.

*) Expression system known, like locally managed Yeast or 'whatever'

*) Selection itself is reliable, UV florescence or some other property

Questions:

Q1) How many labor hours is typical, no paperwork time, just wet lab including end game verification ?

Q2) Whats the success rate of a new germ line emerging in percent ?

Q3) How often is a success rate wrong ultimately. not that the Gene did do what is expected, but some 'other' flaw made it not really get expressed ?

Q4) Is sequencing the believed usable final modified colony invariably reliable enough to make Q3 irrelevant ?

Is Q2) Closer to what ? 30% 80 %

For Q1) I mean hands on time with endless revisits to the plates/wells, dishes, tubes, etc. and so on, not including incubation times, centerfuge, etc.

Thanks in advance of anyone who answers. As people like me want to make this technology, not science, this is a big thing hugely.

Thanks!

Daniel B Kolis

my ref: nafl, 31 Aug 2022, diybio

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