FAQ: Do you have any recommendations for the purification of single-stranded DNA (ssDNA) when using the Monarch® Spin High-Capacity DNA Cleanup Kit (NEB #T1135)?

The Monarch Spin High-Capacity DNA Cleanup Kit (100 μg) can be used to purify ssDNA. ssDNA binds less tightly than dsDNA, and the strength of binding will be affected by length, sequence, and the formation of any secondary structure. For ssDNA < 100 nt, we recommend using the Oligonucleotide Cleanup Protocol provided with the kit, which is optimized to recover shorter DNA. While the Monarch Spin PCR & DNA Cleanup Kit (100 μg) will work well with ssDNA, it is difficult to predict the recovery efficiency of ssDNA. We, therefore, recommend a trial recovery with the DNA of interest before using your entire sample.