Are there any special considerations that must be taken into account when performing AMPure XP purification on libraries prepared with the ThruPLEX DNA-Seq Kit?

Yes, we suggest using a 1:1 bead to sample ratio. Additionally, a freshly prepared solution of 80% must be used in all washing steps of the protocol. Please refer to the ThruPLEX DNA-Seq Kit User Manual for detailed instructions on AMPure XP purification of ThruPLEX DNA-Seq libraries for next-generation sequencing.DNA-seq [...]

Is it necessary to perform quantification of ThruPLEX DNA-Seq libraries before and after the AMPure XP cleanup step?

No, it is only necessary to quantify your individual or pooled libraries prior to sequencing. If you choose to quantify your libraries after the Library Amplification Reaction, it is recommended to do a final quantification of the purified libraries prior to sequencing because AMPure XP purification can result in loss of DNA.DNA-seq [...]

When should library quantification be performed for ThruPLEX DNA-Seq Kit?

Quantification must be performed prior to sequencing. It is normally done after the AMPure XP purification step, but libraries can also be quantified immediately following the Library Amplification Reaction to normalize samples prior to pooling. If yield is of concern, the quantification method used must not require purification of the library. Please refer to the ThruPLEX DNA-Seq Kit User Manual [...]

What is the expected yield for the amplified library from the ThruPLEX DNA-Seq Kit?

The amount of amplified library can range from 100 ng to 1 µg depending upon many variables, including sample type, fragmentation size, and thermal cycler used. When starting with Covaris-fragmented reference DNA with an average size of 200 bp (using the recommended number of amplification cycles in the ThruPLEX DNA-Seq Kit User Manual), typical yields range from 300 to 700 ng. DNA-seq [...]

Can I perform additional amplification of ThruPLEX DNA-Seq libraries after they are purified with AMPure XP beads?

No.DNA-seq [...]