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  3. Biology (-tech,-chem,micro-)

stranded

Russian translation: см.

09:33 Nov 20, 2017
English to Russian translations [PRO]
Science - Biology (-tech,-chem,micro-)
English term or phrase: stranded
After collecting the RNA, it was further purified using an RNeasy mini kit (Qiagen) with an on- column DNase I digestion.
Libraries for Illumina sequencing were generated via the Illumina TruSEQ ***stranded*** mRNA prep kit. Samples were run in a single lane on an Illumina HiSEQ 2000 sequencer with a single read flow cell using 1 x 50-bp reads and a 6-cycle index read. Reads were mapped to the hg19 reference genome using Tophat2 v.2.0.6 with custom settings including the setting of -library-type fr- firstrand to appropriately account for the ***stranded nature*** of the protocol. HTSeq v.0.6.1 was used to obtain read counts over annotated genes and differentially expressed genes were called by DESeq v.1.10.1 with a padj value of less than 0.01.
Counts were normalized for GSEA using the limma voom function. Expression data for the I-BET151 comparison were downloaded from ArrayExpress (https://www.ebi.ac.uk/arrayexpress, accession E-MTAB-774) and processed files used as is. Gene lists were submitted to the DAVID web server (http://david.abcc.ncifcrf.gov) for functional annotation.
Arkadii Marchenko
Ukraine
Local time: 11:07
Russian translation:см.
Explanation:
В первом случае возможно следует оставить оригинальное название набора реагентов.
Во втором перевел бы смысл примерно так: чтобы учесть соответствующим образом особенности протокола, обеспечивающие специфичность [нуклеотидной] цепи, используемой [в дальнейшем] для секвенирования.
либо иными словами
обеспечивающего специфическое секвенирование определенной цепи кДНК

см. Руководство к киту
Introduction
This protocol explains how to convert the mRNA in total RNA into a library of template
molecules of known strand origin using the reagents provided in the Illumina® TruSeq®
Stranded mRNA Sample Preparation Kits. The library is suitable for subsequent cluster
generation and DNA sequencing.
The first step in the workflow involves purifying the poly-A containing mRNA molecules
using poly-T oligo attached magnetic beads. Following purification, the mRNA is
fragmented into small pieces using divalent cations under elevated temperature. The
cleaved RNA fragments are copied into first strand cDNA using reverse transcriptase and
random primers. Strand specificity is achieved by replacing dTTP with dUTP in the Second
Strand Marking Mix (SMM), followed by second strand cDNA synthesis using DNA
Polymerase I and RNase H. The incorporation of dUTP in second strand synthesis
quenches the second strand during amplification, because the polymerase used in the
assay is not incorporated past this nucleotide. The addition of Actinomycin D to First Stand
Synthesis Act D mix (FSA) prevents spurious DNA-dependent synthesis, while allowing
RNA-dependent synthesis, improving strand specificity. These cDNA fragments then have
the addition of a single 'A' base and subsequent ligation of the adapter. The products are
then purified and enriched with PCR to create the final cDNA library.
Selected response from:

Igor Andreev
Local time: 11:07
Grading comment
4 KudoZ points were awarded for this answer



Summary of answers provided
3см.
Igor Andreev


  

Answers


12 hrs   confidence: Answerer confidence 3/5Answerer confidence 3/5
см.


Explanation:
В первом случае возможно следует оставить оригинальное название набора реагентов.
Во втором перевел бы смысл примерно так: чтобы учесть соответствующим образом особенности протокола, обеспечивающие специфичность [нуклеотидной] цепи, используемой [в дальнейшем] для секвенирования.
либо иными словами
обеспечивающего специфическое секвенирование определенной цепи кДНК

см. Руководство к киту
Introduction
This protocol explains how to convert the mRNA in total RNA into a library of template
molecules of known strand origin using the reagents provided in the Illumina® TruSeq®
Stranded mRNA Sample Preparation Kits. The library is suitable for subsequent cluster
generation and DNA sequencing.
The first step in the workflow involves purifying the poly-A containing mRNA molecules
using poly-T oligo attached magnetic beads. Following purification, the mRNA is
fragmented into small pieces using divalent cations under elevated temperature. The
cleaved RNA fragments are copied into first strand cDNA using reverse transcriptase and
random primers. Strand specificity is achieved by replacing dTTP with dUTP in the Second
Strand Marking Mix (SMM), followed by second strand cDNA synthesis using DNA
Polymerase I and RNase H. The incorporation of dUTP in second strand synthesis
quenches the second strand during amplification, because the polymerase used in the
assay is not incorporated past this nucleotide. The addition of Actinomycin D to First Stand
Synthesis Act D mix (FSA) prevents spurious DNA-dependent synthesis, while allowing
RNA-dependent synthesis, improving strand specificity. These cDNA fragments then have
the addition of a single 'A' base and subsequent ligation of the adapter. The products are
then purified and enriched with PCR to create the final cDNA library.

Igor Andreev
Local time: 11:07
Specializes in field
Native speaker of: Native in RussianRussian
PRO pts in category: 1427
Notes to answerer
Asker: С первым случаем все понятно, я выделение сделал, чтобы видна была связь протокола и назначения набора.

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