Hi everyone !
Recently I try to characterize my soil sample with ITS2 region sequencing.
I have used primer 5.8S-Fun/ITS4-Fun from paper Accurate Estimation of Fungal Diversity and Abundance through Improved Lineage-Specific Primers Optimized for Illumina Amplicon Sequencing
which is also used in Fungal ITS analysis tutorial.
The gelelectrophoresis result is a little disappointed since the amplicon length is at least 500bp so it can be sequenced under illumina PE250 strategy.
Which primer should I choose?