Bioanalyzer 2100 dna 1000 kit

PolyA-RNA was isolated from 25 μg of total RNA using the MicroPoly(A) Purist kit (Ambion, USA). Total PolyA-RNA was used to generate whole transcriptome libraries for sequencing on the SOLiD 5500XL platform following the manufacturer’s recommendations (Life Technologies; CA, USA). Amplified cDNA quality was analyzed using the Bioanalyzer 2100 DNA 1000 kit (Agilent Technologies; Spain) and quantified using the Qubit 2.0 Fluorometer (Invitrogen; UK). Whole transcriptome libraries were used to make SOLiD templated beads following the SOLiD templated bead preparation guide. This protocol consisted of an RNA enrichment and chemical modification step, followed by a clonal amplification step. Bead quality was estimated based on work flow analysis parameters. The samples were sequenced using the 50625 paired-end protocol, generating 115 nt sequences consisting of 75 nt plus 35 nt (Paired-End) + 5 nt (Barcode). Quality data was measured using software parameters of the SOLiD Experimental System.

PolyA-RNA was isolated form 25 micrograms of total RNA using the MicroPoly(A) Purist kit (Ambion, USA). Total PolyA-RNA samples were used to generate whole transcriptome libraries for sequencing on the SOLiD 5500XL platform, following the manufacturer's recommendation (Life Technologies, CA). No RNA-spike in controls was used. Amplified cDNA quality was analyzed by the Bioanalyzer 2100 DNA 1000 kit (Agilent Technologies, Spain) and quantified using the Qubit 2.0 Fluorometer (Invitrogen, UK). The whole transcriptome libraries were used for making SOLiD templated beads following the SOLiD Templated Bead Preparation guide. Bead quality was estimated based on WFA (workflow analysis) parameters. The samples were sequenced using the 50625 paired-end protocol, generating 75 nt+35 nt (Paired-End)+5 nt (Barcode) sequences. Quality data were measured using software SETS parameters (SOLiD Experimental Tracking System).

TruSeq Stranded mRNA LT Sample Prep Kit (Illumina,USA,Cat.No.: RS-122-2101); AgencourtAMPure XP (BECKMAN COULTER, USA,Cat.No.:A63881); QubitRNA Assay Kit (LifeTechnologies, USA,Cat.No.:Q32852); Bioanalyzer 2100 RNA-6000 Nano Kit (Aglient, USA,Cat.No.:5067–1511); Bioanalyzer 2100 DNA-1000 Kit (Aglient, USA, Cat.No.:5067–1504); SuperScript II Reverse Transcriptase (Invitroge,USA,Cat.No.: 18064014)

TruSeq Stranded Total RNA with Ribo-Zero Gold (Illumina, United States); AgencourtAMPure XP (BECKMAN COULTER, United States); Agencourt RNAClean XP(BECKMAN COULTER, United States); QubitRNA Assay Kit (Life Technologies, United States); QubitdsDNA Assay Kit (Life Technologies, United States); Bioanalyzer 2100 RNA-6000 Nano Kit (Aglient, United States); Bioanalyzer 2100 DNA-1000 Kit (Agilent, United States); SuperScript II Reverse Transcriptase (Invitrogen, United States); miRNA first strand cDNA synthesis kit (Accurate Biology, China); TRIpure Reagent Total RNA Extraction Reagent (Bioteke Corporation, China); primer (Tsingke Biotechnology Co., China).

The RNA samples were isolated using a MicroPoly(A) Purist Kit (Ambion, USA). The total polyA-RNA samples were used to generate whole transcriptome libraries that were sequenced on a SOLiD 5500XL platform as per the manufacturer’s recommendations (Life Technologies, CA). The amplified cDNA quality was analyzed using the Bioanalyzer 2100 DNA 1000 kit (Agilent Technologies, Spain), and the cDNA was quantified using the Qubit 2.0 Fluorometer (Invitrogen, UK). Whole transcriptome libraries were used to generate SOLiD templated beads by following the SOLiD Templated Bead Preparation guide. Bead quality was estimated based on WFA (workflow analysis) parameters. The samples were sequenced using the 50625 paired-end protocol, which generated 75 nt+35 nt (Paired-End) +5 nt (Barcode) sequences. Quality data were measured using the SETS software parameters (SOLiD Experimental Tracking System).

Poly(A) RNA samples were isolated from 25 µg of total RNA using the MicroPoly(A) Purist kit (Ambion, USA). SOLiD 5500 XL platform was used for sequencing whole transcriptome libraries generated from total Poly(A) RNA samples, following the manufacturer's instructions (Life Technologies, CA, USA). No RNA-spike in CNTs was used. Amplified cDNA quality was analyzed using the Bioanalyzer 2100 DNA 1000 kit (Agilent Technologies, Spain), and quantified using the Qubit 2.0 Fluorometer (Invitrogen, UK). The whole transcriptome libraries were used for making SOLiD-templated beads following the SOLiD System Templated Bead Preparation guidelines. This protocol comprised a clonal amplification step following an enrichment and chemical modification process. Bead quality was estimated based on workflow analysis parameters. The samples were sequenced using the 50625 paired-end protocol, generating 75 nt + 35 nt (paired-end) + 5 nt (barcode) sequences. Quality data was measured using SOLiD Experimental Tracking Software parameters.