| Library |
| Library name |
Construction protocol |
Strategy |
Source |
Selection |
Layout |
|
(1) Oligo (dT) enriched mRNA containing polyA;(2) Reverse transcription of mRNA into cDNA using SMARTer PCR cDNA Synthesis Kit;(3) PCR amplifies and enriches the synthesized cDNA, and determines the optimal PCR conditions through loop optimization;(4) Take some cDNA and use BluePippin for fragment screening, enrich fragments larger than 4Kb, and perform large-scale PCR on the screened fragments to obtain sufficient total cDNA;(5) Full length cDNA was used for damage repair, end repair, and SMRT dumbbell junction connection to construct an equimolar mixed library of non-screened fragments and fragments larger than 4Kb;(6) Nucleic acid exonuclease digestion to remove the sequence of unconnected connectors at both ends of cDNA;(7) Finally, a complete SMRT bell library was formed by binding primers and DNA polymerase. |
ISO-Seq |
TRANSCRIPTOMIC |
PCR |
SINGLE
|
|
