Re: [DIYbio] Re: Bacterial Chimerae or making photosynthetic Bacillus Subtilis

On Wed, Nov 20, 2013 at 2:33 PM, Mega [Andreas Stuermer]
<masterstorm123@gmail.com> wrote:
> Maybe I need to get a microinjector :D
>
> Will that work with such small cells?

It might work, but you'd need smaller needles than for eukaryotes for
sure. It would probably be an order or three of magnitude harder :P

Microinjection on big cells would probably take quite some time, I'd
think at least a year of studying and finding/building apparatus, if
not 2 3 or 4 years!

On the other hand, I think PEG is a much better first try. ;)

I've made hybridomas before, was given the cultured myeloma cells in
suspension (immortal cancer B cell line), and had to slice and dice
and extract from a mouse the spleen and then rinse and smash it to get
the white blood cells out. Then after counting the ratio of B cells to
other WBCs, used overall count to calculate the B cell concentration.
Then we knew how much of culture A and culture B to mix, to get the
desired ratio of myeloma to antibody-producing B cells. That ratio
could vary, this paper says spleen cell to myeloma cell ratio is 3:1,
but can vary from 1:1 to 10:1. (pg 393)
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2596021/pdf/yjbm00116-0074.pdf


I did some searching, found this but can't get it, will let you know
if I can get it though:
http://link.springer.com/chapter/10.1007/978-1-4684-4142-0_7

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