Topic: cloning
Stance: CON
Aspect: time-limiting tactics
However , in our hands , we found that these methods were too limited by their own inherent limitations : 1 ) Time limits for subclonings can vary widely among strains ; 2 ) Cloning into plasmids or bacteriophage derivatives is often difficult because most phages recognize only one copy of DNA at any given point in the genome [ 11 ] ; 3 ) Restriction enzymes play an important role in bacterial reproduction [ 12 - 14 ] , which makes traditional restriction enzyme based systems less amenable than previously thought to accommodate multiple gene insertions and/or deletions without loss of protein function ; 4 ) Because many genes encode proteins having different biological properties when expressed in E. coli , standard restriction-dependent cloning approaches using lambda library technology are slow and labor intensive [ 15 - 17 ] .