3 replies to this topic

[jungskwon98]

I am thinking of doing Double linker ligation into a vector.  Here is the scheme.

• Anneal Linker 1(phospolilated 1a oligo + phospolilated 1b oligo)
  
• 
  Anneal Linker 2(phospolilated 2a oligo + phospolilated 2b oligo)
  
• 
  Ligated Linker 1+2 with T4, gel purify correct size
  
• 
  Digest and CIP vector
  
• 
  Ligate Vector with Linker (1+2)

Do you guys think this will work???

Due to no digestion site and price(of oligo synthesis) I am looking for alternatives.

Thank you in advance for your help!!!!

[bob1]

It should work, that was how cloning was done before the advent of PCR generated restriction sites.

[jungskwon98]

Sorry for the late reply but Thank you bob1.

I have already went ahead with the protocol in mind will update if someone needs to do this.

[jungskwon98]

bob1, on 03 May 2012 - 12:57 PM, said:

It should work, that was how cloning was done before the advent of PCR generated restriction sites.

Sorry for the late reply but Thank you bob1.

I have already went ahead with the protocol in mind will update if someone needs to do this.