The sequencing library was prepared according to the Iso‐Seq protocol as described by Pacific Biosciences (P/N100‐377‐100‐05 and P/N100‐377‐100‐04). The SMARTer PCR cDNA Synthesis Kit was used to synthesize cDNA from the same RNA samples used for Illumina sequencing. After 23 cycles of PCR amplification, products were size selected using the BluePippin Size Selection System with the following bins for each sample: 1–2, 2–3 and 3–10 kb. The amplified cDNA products were used to generate SMRTbell Template libraries according to the Iso‐Seq protocol. Libraries were prepared for sequencing by annealing a sequencing primer and adding polymerase to the primer‐annealed template. The polymerase‐bound template was bound to MagBeads and sequencing was performed on a PacBio RSII instrument.
Smarter pcr cdna synthesis kit
Manufactured by Illumina
Sourced in China
The SMARTer PCR cDNA Synthesis Kit is a laboratory equipment product that enables the synthesis of complementary DNA (cDNA) from RNA samples. The kit provides a method for generating high-quality cDNA libraries from small amounts of input RNA.
Automatically generated - may contain errors
Lab products found in correlation
Promethion, by Oxford Nanopore Technologies (2 mentions)
Direct cdna sequencing kit, by Oxford Nanopore Technologies (2 mentions)
Gridion x5, by Oxford Nanopore Technologies (2 mentions)
Ligation sequencing kit, by Oxford Nanopore Technologies (2 mentions)
Novaseq platform, by Illumina (2 mentions)
Total rna extraction kit, by Takara Bio (1 mentions)
Rna seq, by Pacific Biosciences (1 mentions)
Hiseq x ten platform, by Illumina (1 mentions)
2100 bioanalyzer rna nanochip, by Agilent Technologies (1 mentions)
Rnaeasy plant mini kit, by Qiagen (1 mentions)
Rnaprep pure plant plus kit, by Tiangen Biotech (1 mentions)
Hiseq 2500 sequencing platform, by Illumina (1 mentions)
Pe150, by Novogene (1 mentions)
Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions)
6 protocols using smarter pcr cdna synthesis kit
1
Long-Read Transcriptome Sequencing
2
Integrative Transcriptome Analysis of Marchantia
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We performed RNA-seq analysis using the Illumina RNA-seq (separate samples) and PacBio full-length transcript (mixed samples) sequencing methods to improve the predictive ability of gene models. Six different tissues from M. polymorpha, including seeds, roots, stems, leaves, flowers, and pods, were collected and frozen in liquid nitrogen. Total RNA was extracted using a Takara Total RNA Extraction Kit (Takara, Dalian, China). Transcriptome libraries were constructed using a NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) according to the manufacturer’s procedures. Then, 150-bp paired-end sequencing was performed on the Illumina HiSeq X Ten platform. For PacBio sequencing, cDNA was synthesized from the mixed RNA samples used for Illumina sequencing using a SMARTer PCR cDNA Synthesis Kit. After PCR amplification, the products were sequenced on the PacBio RSII platform.
3
Full-length cDNA Library Synthesis and Sequencing
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Total RNAs were extracted using RNA prep Pure Plant plus Kit (Tiangen Biotech (Beijing) Co., Ltd., Beijing, China) and purified with the RNA easy Plant Mini Kit (Qiagen, Valencia, CA, USA). RNA quality was verified using a 2100 Bioanalyzer RNA Nanochip (Agilent, Santa Clara, CA, USA), and quantified using NanoDrop ND-1000 Spectrophotometer (Nano-Drop, Wilmington, DE, USA).
For full-length cDNA library, cDNA was synthesized using SMARTer PCR cDNA Synthesis Kit, and optimized for PCR amplification. The fragments for large-scale PCR were performed using magnetic beads to obtain sufficient total cDNA. The complete SMRT bell library was constructed with using a SMARTer PCR cDNA Synthesis Kit and assembly was performed on the PacBio Sequel platform, the second-generation sequencing and assembly was implemented on the Hiseq 2500 sequencing platform (Illumina) with PE150 by Novogene Co., Ltd. (Beijing, China).
For full-length cDNA library, cDNA was synthesized using SMARTer PCR cDNA Synthesis Kit, and optimized for PCR amplification. The fragments for large-scale PCR were performed using magnetic beads to obtain sufficient total cDNA. The complete SMRT bell library was constructed with using a SMARTer PCR cDNA Synthesis Kit and assembly was performed on the PacBio Sequel platform, the second-generation sequencing and assembly was implemented on the Hiseq 2500 sequencing platform (Illumina) with PE150 by Novogene Co., Ltd. (Beijing, China).
4
Multi-platform Transcriptome Sequencing
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For Illumina sequencing, cDNA libraries (with three biological replicates) were constructed using the NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs, Beverly, MA, USA) following the manufacturer’s protocol. The libraries were sequenced on an Illumina NovaSeq platform, and paired-end reads were generated.
For PacBio sequencing, cDNA from the same RNA samples used for Illumina sequencing was synthesised using the SMARTer PCR cDNA Synthesis Kit. After PCR amplification, products were sequenced on the PacBio Sequel platform.
For ONT sequencing, cDNA-PCR libraries were built using the Ligation Sequencing Kit (SQK-LSK109) and sequenced on the Nanopore PromethION platform. Direct-cDNA libraries were built using the Direct cDNA Sequencing Kit (SQK-DCS108) and sequenced on the Nanopore GridION X5 platform.
For PacBio sequencing, cDNA from the same RNA samples used for Illumina sequencing was synthesised using the SMARTer PCR cDNA Synthesis Kit. After PCR amplification, products were sequenced on the PacBio Sequel platform.
For ONT sequencing, cDNA-PCR libraries were built using the Ligation Sequencing Kit (SQK-LSK109) and sequenced on the Nanopore PromethION platform. Direct-cDNA libraries were built using the Direct cDNA Sequencing Kit (SQK-DCS108) and sequenced on the Nanopore GridION X5 platform.
5
Multiplatform Transcriptome Sequencing
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For Illumina sequencing, cDNA libraries (with three biological replicates) were constructed using the NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs, Beverly, MA, USA) following the manufacturer's protocol. The libraries were sequenced on an Illumina NovaSeq platform, and paired-end reads were generated.
For PacBio sequencing, cDNA from the same RNA samples used for Illumina sequencing was synthesised using the SMARTer PCR cDNA Synthesis Kit. After PCR ampli cation, products were sequenced on the PacBio Sequel platform.
For ONT sequencing, cDNA-PCR libraries were built using the Ligation Sequencing Kit (SQK-LSK109) and sequenced on the Nanopore PromethION platform. Direct-cDNA libraries were built using the Direct cDNA Sequencing Kit (SQK-DCS108) and sequenced on the Nanopore GridION X5 platform.
For PacBio sequencing, cDNA from the same RNA samples used for Illumina sequencing was synthesised using the SMARTer PCR cDNA Synthesis Kit. After PCR ampli cation, products were sequenced on the PacBio Sequel platform.
For ONT sequencing, cDNA-PCR libraries were built using the Ligation Sequencing Kit (SQK-LSK109) and sequenced on the Nanopore PromethION platform. Direct-cDNA libraries were built using the Direct cDNA Sequencing Kit (SQK-DCS108) and sequenced on the Nanopore GridION X5 platform.
6
PacBio Sequencing of Fruit cDNA
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The SMARTer PCR cDNA Synthesis Kit was used to synthesize cDNA from the same RNA samples used for Illumina sequencing. The cDNA was synthesized using mixed RNAs from five fruit developmental stages for each mesocarp and seed sample. After the PCR amplification, quality control analysis, and purification, fulllength cDNA fragments were obtained with the BluePippin Size Selection System protocol for the construction of each mesocarp and seed cDNA library (1-6 kb). Selected full-length cDNA was ligated to the SMRT bell hairpin loop. A Qubit 2.0 fluorometer was then used to determine the cDNA library concentration. The quality of the cDNA library was assessed with the Agilent 2100 Bioanalyzer. Finally, each SMRT cell for the mesocarp and seed was sequenced with the PacBio Sequel instrument (Pacific Bioscience, Menlo Park, CA, USA).
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