x********u
发帖数: 430 | 1 I am going to clone an operon with two genes, the total length is about 4500 bp. Is it possible that I clone this big fragment all at once and get the
right sequence? I am using the dual-promoter system and E.coli host strain. Many
thanks for your help. |
H****N
发帖数: 997 | 2 This should be a piece of cake |
x********u
发帖数: 430 | 3 Thanks. I will try that.
Some people suggested me to select some unique RE sites and clone part of
the fragment first, then comes to the other part of the fragment. I dno't
think this is a good choice. |
n********k
发帖数: 2818 | 4 I concur with HENBEN...BTW, if u come here to do brain sections for a week
or two, I would do as many of this
kind cloning as u like:))...be a little careful with if you are building ur own bicistro system though...but I am
assuming u are using commercial vector, then it shall be easy...
【在 x********u 的大作中提到】
: Thanks. I will try that.
: Some people suggested me to select some unique RE sites and clone part of
: the fragment first, then comes to the other part of the fragment. I dno't
: think this is a good choice.
|
X***n
发帖数: 366 | 5 Try SLIC.
Nature Methods - 4, 251 - 256 (2007)
Published online: 11 February 2007; | doi:10.1038/nmeth1010
Harnessing homologous recombination in vitro to generate recombinant DNA via
SLIC
Mamie Z Li & Stephen J Elledge
4500 bp. Is it possible that I clone this big fragment all at once and get
the
. Many
【在 x********u 的大作中提到】
: I am going to clone an operon with two genes, the total length is about 4500 bp. Is it possible that I clone this big fragment all at once and get the
: right sequence? I am using the dual-promoter system and E.coli host strain. Many
: thanks for your help.
|
x********u
发帖数: 430 | |
