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