I am planing to do the trascriptome analysis using PacBio technology.
After searching and reading the protocols, I know there are two methods used by people to generate full-length cDNA library for PacBio sequencing: 1. using "SuperScript Full-Length cDNA Library Construction Kit" from Invitrogen. The advantage of this method is that in this kit, 5'Cap specific Ab is used for selection of full-length of cDNA. But, unfortunately, this kit is discontinued for unknown reason (Anyone know the reason?)
2. using Clontech SMART Kit, which is also recommended by PacBio company.
Now, my 1st question is: Are there any better methods for full-length cDNA synthesis available and compatible with PacBio besides those two I mentioned above?
Such as, dUTP method (http://nar.oxfordjournals.org/content/37/18/e123.long) which is almost standard method for Illumina sequencing.
Or, "first-strand cDNA library" method (Sequencing of first-strand cDNA library reveals full-length transcriptomes: http://www.nature.com/ncomms/2015/15...comms7002.html).
My 2nd Q: Whether these two methods also can be used for PacBio platform?
Any input/thought will be appreciated!
After searching and reading the protocols, I know there are two methods used by people to generate full-length cDNA library for PacBio sequencing: 1. using "SuperScript Full-Length cDNA Library Construction Kit" from Invitrogen. The advantage of this method is that in this kit, 5'Cap specific Ab is used for selection of full-length of cDNA. But, unfortunately, this kit is discontinued for unknown reason (Anyone know the reason?)
2. using Clontech SMART Kit, which is also recommended by PacBio company.
Now, my 1st question is: Are there any better methods for full-length cDNA synthesis available and compatible with PacBio besides those two I mentioned above?
Such as, dUTP method (http://nar.oxfordjournals.org/content/37/18/e123.long) which is almost standard method for Illumina sequencing.
Or, "first-strand cDNA library" method (Sequencing of first-strand cDNA library reveals full-length transcriptomes: http://www.nature.com/ncomms/2015/15...comms7002.html).
My 2nd Q: Whether these two methods also can be used for PacBio platform?
Any input/thought will be appreciated!
Comment