Hi everyone,
I need to amplify selected regions from my RNA samples and resequence with MiSeq.
I am unclear to what is the best approach:
1. Use the Illumina metagenomic 16s sequencing library kit, design PCR primers with the Illumina adapter sequences attached as part of the primers. Pool the amplicons. Then I use the Nextera index kit to index the samples.
or
2. Perform my own PCR independent of Illumina kits, purify the PCR fragments and then add adapter and index sequences.
Advice on multiplexing PCR software is also highly appreciated!
Regards,
Dessi
I need to amplify selected regions from my RNA samples and resequence with MiSeq.
I am unclear to what is the best approach:
1. Use the Illumina metagenomic 16s sequencing library kit, design PCR primers with the Illumina adapter sequences attached as part of the primers. Pool the amplicons. Then I use the Nextera index kit to index the samples.
or
2. Perform my own PCR independent of Illumina kits, purify the PCR fragments and then add adapter and index sequences.
Advice on multiplexing PCR software is also highly appreciated!
Regards,
Dessi
Comment