TY - CHAP
T1 - Insertion and deletion mutagenesis by overlap extension PCR
AU - Lee, Jehan
AU - Shin, Myeong Kyun
AU - Ryu, Dong Kyun
AU - Kim, Seahee
AU - Ryu, Wang Shick
PY - 2010
Y1 - 2010
N2 - Mutagenesis by the overlap extension PCR has become a standard method of creating mutations including substitutions, insertions, and deletions. Nonetheless, the established overlap PCR mutagenesis is limited in many respects. In particular, it has been difficult to make an insertion larger than 30 nt, since all sequence alterations must be embedded within the primer. Here, we describe a rapid and efficient method for creating insertions or deletions of any length at any position in a DNA molecule. This method is generally applicable, and therefore represents a significant improvement to the now widely used overlap extension PCR method.
AB - Mutagenesis by the overlap extension PCR has become a standard method of creating mutations including substitutions, insertions, and deletions. Nonetheless, the established overlap PCR mutagenesis is limited in many respects. In particular, it has been difficult to make an insertion larger than 30 nt, since all sequence alterations must be embedded within the primer. Here, we describe a rapid and efficient method for creating insertions or deletions of any length at any position in a DNA molecule. This method is generally applicable, and therefore represents a significant improvement to the now widely used overlap extension PCR method.
KW - Chimeric primers
KW - Deletion
KW - Insertion
KW - Overlap extension PCR
KW - Site-directed mutagenesis
UR - http://www.scopus.com/inward/record.url?scp=79952752445&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79952752445&partnerID=8YFLogxK
U2 - 10.1007/978-1-60761-652-8_10
DO - 10.1007/978-1-60761-652-8_10
M3 - Chapter
C2 - 20676981
AN - SCOPUS:79952752445
SN - 9781607616511
T3 - Methods in Molecular Biology
SP - 137
EP - 146
BT - In Vitro Mutagenesis Protocols
A2 - Jeff, Braman
ER -