454 GS Junior sequencing of SAMD00018025
The host-plant chloroplast large-subunit of ribulose-1,5-bisphosphate caboxylase/oxygenase (rbcL) region were sequenced using a tag-encoded, massively parallel pyrosequencing analysis. For each root sample, a 0.5-kb rbcL gene fragment was amplified using the forward primer rbcL_F3 (5??-AYT CCC AAC CAT TYA TGC G-3??) fused with the 454 pyrosequencing Adaptor A (5??-CCA TCT CAT CCC TGC GTG TCT CCG ACT CAG-3??) and the 8-mer molecular ID  of each sample, and the reverse primer rbcL_R4 (5??-CAT ATG CCA AAC RTG AAT ACC-3??) fused with the 454 Adaptor B (5??-CCT ATC CCC TGT GTG CCT TGG CAG TCT CAG-3??). The PCR was conducted based on the experimental conditions detailed elsewhere. The rbcL amplicons were subjected to pyrosequencing. The PCR products were pooled and subjected to a purification step using ExoSAP-IT (GE Healthcare, UK) and QIAquick PCR Purification Kit (QIAGEN). The 454 pyrosequencing was performed with a GS Junior sequencer (Roche) according to the manufacturers?f instructions.