Pooling PacBio libraries
Hello all, glad to join the seqanswers community! I am a new PacBio user and hoping to use this forum to get some advice on pooling and sequencing libraries.
The project I am currently planning involves the sequencing of 5 HLA loci. The PCR for HLA-DRB1 produces two products of about 2.5kb and 5kb (which must be pooled together), while the rest are single products of between 3.4kb and 5.8kb.
I am concerned about the pooling of a smaller 2.5kb product with 5+kb ones. If anyone has experience of this protocol or similar, and achieved sufficient coverage, it would be great to chat! I couldn't find a current thread on this subject so apologies if I've missed one.
Thanks in advance,
Hannah
The project I am currently planning involves the sequencing of 5 HLA loci. The PCR for HLA-DRB1 produces two products of about 2.5kb and 5kb (which must be pooled together), while the rest are single products of between 3.4kb and 5.8kb.
I am concerned about the pooling of a smaller 2.5kb product with 5+kb ones. If anyone has experience of this protocol or similar, and achieved sufficient coverage, it would be great to chat! I couldn't find a current thread on this subject so apologies if I've missed one.
Thanks in advance,
Hannah