Differences

This shows you the differences between two versions of the page.

Link to this comparison view

Both sides previous revision Previous revision
interactome:experiments:20191210-3 [2019/12/11 12:46]
dan [Table]
interactome:experiments:20191210-3 [2019/12/11 12:46] (current)
dan
Line 1: Line 1:
 ## Personal Interactome ## Personal Interactome
-## One-by-one pENTRY clone generation from HEK293Ta cDNA sample+# One-by-one pENTRY clone generation from HEK293Ta cDNA sample
  
 == BP cloning== == BP cloning==
  
  
-#### Material+=== Material ​===
   * pExpression plasmid sample (eg. pAD-ORF) or PCR product with attB sequencing flanking the desired gene fragment.   * pExpression plasmid sample (eg. pAD-ORF) or PCR product with attB sequencing flanking the desired gene fragment.
   * pDONR plasmid sample (pDONR221, pDONR223, etc.)   * pDONR plasmid sample (pDONR221, pDONR223, etc.)
Line 28: Line 28:
  
  
-#### Protocol+=== Protocol===
  
-====1  Substrate preparation====+==1  Substrate preparation==
   * Prepare 6-60ng/ul solution of attB PCR sample/ plasmids.   * Prepare 6-60ng/ul solution of attB PCR sample/ plasmids.
  
Line 36: Line 36:
   * Add 2ul of the DNA as follows into one PCR plate.   * Add 2ul of the DNA as follows into one PCR plate.
     
-====2  BP reaction====+==2  BP reaction==
  
   *Prepare Ice with a tube labled "​BP"​   *Prepare Ice with a tube labled "​BP"​
Line 56: Line 56:
  
   
-====3  Transformation====+==3  Transformation==
   *Prepare the reaction mix and competent cells on ice.   *Prepare the reaction mix and competent cells on ice.
   *For each sample, add 10µl of homebrew competent NEB5-α cells.   *For each sample, add 10µl of homebrew competent NEB5-α cells.
  • interactome/experiments/20191210-3.1576035973.txt.gz
  • Last modified: 2019/12/11 12:46
  • by dan