TY - JOUR
T1 - Polymerase chain reaction-based gene removal from plasmids
AU - Krishnamurthy, Vishnu Vardhan
AU - Khamo, John S.
AU - Cho, Ellen
AU - Schornak, Cara
AU - Zhang, Kai
N1 - KZ thanks the funding support from University of Illinois at Urbana-Champaign (UIUC), United States . We thank Prof. Joseph Gall at the Carnegie Institution for Science for providing the pCRII plasmid and Prof. XL Nan in the Oregon Health and Science University for providing the ERK-GFP plasmid. We thank Dr. Sandra McMasters from the cell media facility of UIUC for providing DH5α competent cells and for her insightful comments on our experiments.
PY - 2015/9/1
Y1 - 2015/9/1
N2 - This data article contains supplementary figures and methods to the research article entitled, "Multiplex gene removal by two-step polymerase chain reactions" (Krishnamurthy et al., Anal. Biochem., 2015, doi:http://dx.doi.org/10.1016/j.ab.2015.03.033), which presents a restriction-enzyme free method to remove multiple DNA segments from plasmids. Restriction-free cloning methods have dramatically improved the flexibility and speed of genetic manipulation compared to conventional assays based on restriction enzyme digestion (Lale and Valla, 2014. DNA Cloning and Assembly Methods, vol. 1116). Here, we show the basic scheme and characterize the success rate for single and multiplex gene removal from plasmids. In addition, we optimize experimental conditions, including the amount of template, multiple primers mixing, and buffers for DpnI treatment, used in the one-pot reaction for multiplex gene removal.
AB - This data article contains supplementary figures and methods to the research article entitled, "Multiplex gene removal by two-step polymerase chain reactions" (Krishnamurthy et al., Anal. Biochem., 2015, doi:http://dx.doi.org/10.1016/j.ab.2015.03.033), which presents a restriction-enzyme free method to remove multiple DNA segments from plasmids. Restriction-free cloning methods have dramatically improved the flexibility and speed of genetic manipulation compared to conventional assays based on restriction enzyme digestion (Lale and Valla, 2014. DNA Cloning and Assembly Methods, vol. 1116). Here, we show the basic scheme and characterize the success rate for single and multiplex gene removal from plasmids. In addition, we optimize experimental conditions, including the amount of template, multiple primers mixing, and buffers for DpnI treatment, used in the one-pot reaction for multiplex gene removal.
KW - Gene removal
KW - Plasmids
KW - Polymerase chain reaction
KW - Restriction-free cloning
UR - https://www.scopus.com/pages/publications/84929989829
UR - https://www.scopus.com/pages/publications/84929989829#tab=citedBy
U2 - 10.1016/j.dib.2015.04.024
DO - 10.1016/j.dib.2015.04.024
M3 - Article
AN - SCOPUS:84929989829
SN - 2352-3409
VL - 4
SP - 75
EP - 82
JO - Data in Brief
JF - Data in Brief
ER -