Developing an insert to protect large DNA molecules during cell lysis.

Abstract

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 S. cerevisiae cells to show that cells could be lysed inside the inverted insert, allowing the DNA to remain at full length in the middle.