Tissues of leaves, stems, roots, and flowers were collected from 30-day-old Zhongzhu 1 as a sample mixture and immediately frozen in liquid nitrogen. Total RNA of this sample was extracted using an E.Z.N.A. Plant RNA Kit (OMEGA Bio-tek, Norcross, GA, USA) according to the manufacturer’s protocol and used for single-molecule sequencing. In brief, approximately 1 μg of total RNA was used to construct an Iso-Seq SMRTBell library after a reverse-transcription reaction using a Clontech SMARTer cDNA Synthesis Kit (Clontech Laboratories, Mountain View, CA, USA) and oligo dT primer to generate full-length cDNA. Then, the library was subjected to single-molecule sequencing using a PacBio Sequel platform.
Clontech smarter cdna synthesis kit
Manufactured by Takara Bio
Sourced in United States
The Clontech SMARTer cDNA Synthesis Kit is a tool for generating high-quality full-length cDNA from small amounts of input RNA. It utilizes the company's proprietary SMART (Switching Mechanism At 5' end of RNA Template) technology to produce cDNA with intact 5' ends.
Automatically generated - may contain errors
Lab products found in correlation
Plant rna kit, by Omega Bio-Tek (1 mentions)
Ampure pb bead, by Pacific Biosciences (1 mentions)
Facsaria sorter, by BD (1 mentions)
Ficoll paque plus gradient, by GE Healthcare (1 mentions)
Rneasy plus micro kit, by Qiagen (1 mentions)
Dna template prep kit 2, by Pacific Biosciences (1 mentions)
Bluepippin size selection system, by Sage Science (1 mentions)
4 protocols using clontech smarter cdna synthesis kit
1
RNA Extraction and Sequencing from Plant Tissues
2
Iso-Seq Transcriptome Profiling of Garlic
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The landrace from Yangxi (China) was selected among the 102 garlic landraces for single-molecule sequencing. Approximately 1 µg of total RNA of developing bulbs was used for reverse transcription (RT) using a Clontech SMARTer cDNA synthesis kit (Clontech Laboratories, Mountain View, CA, USA) and oligo dT primer to generate full-length cDNA. Three RT reactions were run in parallel, and primer IIA from the kit was used for all PCR reactions following RT. The optimal amplification cycle number for generating dsDNA was determined. PCR products were purified with AMPure PB beads (Pacific Biosciences, Menlo Park, CA, USA). The obtained dsDNA was subjected to size selection using the BluePippin System and subsequent re-amplification. Finally, the products were purified and subjected to Iso-Seq SMRTBell library preparation (https://pacbio.secure.force.com/SamplePrep ) to yield three libraries (1–2, 2–3, and 3–6 kb), which were sequenced on the PacBio RSII platform using P6-C4 chemistry.
3
Isolation and Sorting of Immune Cells from JDM Patients
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PBMCs from 4 new-onset JDM patients and 4 healthy pediatric controls were isolated using a Ficoll-Paque PLUS gradient (GE Healthcare) and cryopreserved. Immune cells were stained with CD56 (NCAM-1) BV421, CD3 (UCHT1) PerCP-Cy5.5, CD14 (M5E2) APC-Fire750, and CD19 (SJ25C1) FITC (Biolegend, San Diego, CA) and sorted to high purity using a FACSAria sorter (BD, Franklin Lakes, NJ). Live cells were sorted into three groups: NK cells (CD56+, CD3-, CD14-), B cells (CD19+, CD3-, CD14-), and T cells (CD3+, CD56-, CD14-) (Supplemental Figure 2
4
Long-read cDNA Sequencing Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Using the Clontech SMARTer cDNA Synthesis Kit (TaKaRa, Japan), 5 μg of mixed total RNA was reverse-transcribed into cDNA. The BluePippin Size Selection System (Sage Science, MA) was used for size fractionation and selection. SMRT sequencing libraries were constructed using the Pacific Biosciences DNA Template Prep Kit 2.0. Finally, SMRT cells were sequenced on the Pacific Bioscience RS II platform.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!