Journal: International Journal of Molecular Sciences
Article Title: Molecular Alterations in Spermatozoa of a Family Case Living in the Land of Fires—A First Look at Possible Transgenerational Effects of Pollutants
doi: 10.3390/ijms21186710
Figure Lengend Snippet: DNA binding ability of SNBP obtained from: control 1 ( a ); son ( b ) and father ( c ) analyzed by EMSA on 1% agarose gel. Bands on gel representing the state of pGEM3 plasmid DNA incubated in a w/w ratio with increasing amount of SNBP from samples containing: CP/Hr ( a ); nCP/Hr ( b ); only–H ( c ). Panel a from supplementary material of Lettieri et al., 2020 . L-group: low impact area group; EMSA: electrophoresis mobility shift Assay; SNBP: sperm nuclear basic proteins; CP/Hr: canonical protamines/histones ratio; nCP/Hr: not canonical protamines/histones ratio; only-H: only histones and other basic proteins.
Article Snippet: pGEM3 plasmid (2867 bp) was extracted from transformed Escherichia coli HB 101 cells, using the standard protocol of the QIAGEN Plasmid Midi Purification kit (QIAGEN Plasmid Midi Purification handbook, third edition© 2020, Hilden, Germany), but following the precautions described in Carbone et al., 2012 [ ], in order to obtain high amounts of supercoiled pDNA.
Techniques: Binding Assay, Control, Agarose Gel Electrophoresis, Plasmid Preparation, Incubation, Electrophoresis, Mobility Shift