FAQ: Do you have any recommendations for the purification of single-stranded DNA (ssDNA) when using the Monarch® Spin PCR & DNA Cleanup Kit (NEB #T1130)?
The Monarch Spin PCR & DNA Cleanup Kit (5 μg) can be used to purify ssDNA from enzymatic reactions, as well as for general cleanup to remove potential inhibitors of PCR. 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 < 200 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 (5 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.