Developing an insert to protect large DNA molecules during cell lysis

Determining large structural variations is difficult and time-consuming without long molecules to aid in the genome assembly. One issue is the fragility of large DNA molecules during routine molecular biology techniques. A novel inverted agarose insert was created to protect and make accessing large DNA easier. The inverted agarose insert has the cell solution in the middle and the agarose on the outside rather than the cells embedded in the agarose (plug inserts). Multiple agarose concentrations were tested to determine the best percentage of agarose to create the inverted insert. A proof-of-principle inverted insert was tested with <i>S. cerevisiae</i> cells to show that cells could be lysed inside the inverted insert, allowing the DNA to remain at full length in the middle. A new inverted insert was developed where the cell solution is in the middle, and the outside has a higher gel percent layer. The cells are lysed, and the DNA stays in the middle of the insert in solution. The inverted inserts make it easy to protect and access the DNA when needed. The inverted insert stored a dynamic range of DNA sizes, and the DNA remained intact for extended periods.More DNA remained in the inverted insert compared to the original plug insert.The inverted inserts stored DNA for long periods kept the chromosomes intact, and allowed easy access to the DNA solution.A proof-of-principle inverted insert was tested with <i>S. cerevisiae</i> cells to show that cells could be lysed inside the inverted insert, allowing the DNA to remain at full length in the middle. The inverted insert stored a dynamic range of DNA sizes, and the DNA remained intact for extended periods. More DNA remained in the inverted insert compared to the original plug insert. The inverted inserts stored DNA for long periods kept the chromosomes intact, and allowed easy access to the DNA solution. A proof-of-principle inverted insert was tested with <i>S. cerevisiae</i> cells to show that cells could be lysed inside the inverted insert, allowing the DNA to remain at full length in the middle.