930
edits
Changes
added ncbi primer blast
#Pick 5' and 3' restriction sites that are present in the vector but NOT in the insert
#Check restriction sites either with NEBCutter (http://tools.neb.com/NEBcutter2/index.php)
#If restriction sites are not available look for sites with compatible cohesive ends (http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/compatible_cohesive_overhangs.asp). #For example:**BamHI = BclI = BglII; **EcoRI = MfeI = ApoI**XbaI = AvrII = SpeI = NheI**XhoI = SalI = PspXI
==Design primers==
#Design 3' end of primers with NCBI Primer Blast (http://www.ncbi.nlm.nih.gov/tools/primer-blast) or Primer3Plus (http://www.bioinformatics.nl/cgi-bin/primer3plus/primer3plus.cgi).
#Adjust the 5' end of primers as below:
##If making a fusion with a N-terminal tag:
