Hello guys,
Hi Guys,
I am preparing to use PacBio Isoforms sequencing, to sequence the total human RNA searching for specific transcripts, which are 7.4-7.7kb long, and occurring in a very low frequency (~1%, if not lower) among total RNA. The main goal is to characterise and quantify these specific, low-frequency RNA molecules. I want to kindly ask for your advice as I am not sure of the following points:
1) How deep could the PacBio Iso-Seq detect in terms of number of transcript? I have been searching in vein to find a reference for this!
2) Will it help to reliably detect those rare molecules if I increase the number of SMRTcells?
3) Taking into consideration the length of the target transcripts (7.4-7.7kb), how many full transcript could I retrieve with a reasonable number of passes, say around >=3, in other words, how good is the chance for at least 1/3 of ZMW productivity 1 to get a high quality Read of Insert?
Many thanks
Hi Guys,
I am preparing to use PacBio Isoforms sequencing, to sequence the total human RNA searching for specific transcripts, which are 7.4-7.7kb long, and occurring in a very low frequency (~1%, if not lower) among total RNA. The main goal is to characterise and quantify these specific, low-frequency RNA molecules. I want to kindly ask for your advice as I am not sure of the following points:
1) How deep could the PacBio Iso-Seq detect in terms of number of transcript? I have been searching in vein to find a reference for this!
2) Will it help to reliably detect those rare molecules if I increase the number of SMRTcells?
3) Taking into consideration the length of the target transcripts (7.4-7.7kb), how many full transcript could I retrieve with a reasonable number of passes, say around >=3, in other words, how good is the chance for at least 1/3 of ZMW productivity 1 to get a high quality Read of Insert?
Many thanks
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