[DIYbio] Re: help me setup DIY-BIO home Lab to produce Recombinant Factor VIII

can i put forwarding few viewpoints just as thought provoking possibilities 

for discussing "why we always end up at bacterium as cell line always and
for every single recommended protein"

 

yes guys I accept it's less complex, less expensive and well established 
over decades and as every scientist I too believe in Occam's razor "law of parsimony", 

"when you have two competing theories that make exactly the same predictions, the simpler one is the better." 

but it Shouldn't stop us ideation on even simpler and everlasting approaches as Occam's razor also states 

"Everything should be made as simple as possible, but not simpler." why can't we adopt 

Agrobacterium-mediated Transient Protein Production in Nicotiana as it gives even 

simpler and everlasting approach to producing Protein of our interest, 

let me chalk down few Advantages and disadvantages of Agrobacterium.

 

Advantages:
- technically simple
- yields relatively uncomplicated insertion events (low copy number,minimal rearrangements)
- unlimited size of foreign DNA
- efficient (for most plants)
- adaptable to different cell types, culture procedures (protoplasts,tissue sections, "non-culture" methods)
- transformants are mitotically and meiotically stable

Disadvantages
- host range is limited: not all plants may be susceptible to Agrobacterium
- with susceptible plants, accessible culture/regeneration systems must be adaptable to Agrobacterium-mediated gene transfer

 

pls friends let me know your opinion in this regard

 

Regard 
Sunil Phani


On Saturday, January 17, 2015 at 7:31:14 AM UTC+5:30, Sunil Phani wrote:


can someone please help me setup DIY-BIO home Lab to perform mammalian cell culture to produce  Recombinant Factor VIII protein with  mammalian cells(CHO, HKE 293, HEL, etc) and kindly provide me recombinant dna protocols for identification, isolation , extraction, insertion and expansion .my requirement is to prepare pilot project in small scale DIY-BIO lab which can further be scaled.

if possible provide me vectors, cell line and seedstock. i am new to DIY-BIO hence i need helping hands of veterans like you ppl .. your help be highly appreciated


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