Extraction of high-quality tissue-specific RNA from London plane trees (Platanus acerifolia), permitting the construction of a female inflorescence cDNA library

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Extraction of high-quality tissue-specific RNA from London plane trees (Platanus acerifolia), permitting the construction of a female inflorescence cDNA library

The London airplane tree (Platanus acerifolia Willd.) has world significance as an city landscaping tree and is the topic of genetic-improvement applications for productive sterility, illness and/or insect resistance. Molecular evaluation methods are essential to such applications, however could also be impeded by particular difficulties encountered throughout nucleic acid isolation.

An in depth RNA isolation and purification protocol, primarily based on established cetyltrimethyl-ammonium bromide (CTAB) extraction methods mixed with extra purification steps utilizing butanol and the ionic detergent CTAB, which overcomes these issues within the woody species P. acerifolia, was carried out. Briefly, phenolic compounds are certain to soluble polyvinylpyrrolidone after which separated out by LiCl precipitation of the RNA. Subsequently, protein- and carbohydrate-contaminants are eliminated by chloroform partitioning adopted by LiCl-mediated precipitation.

The ensuing isolates of RNA had been discovered to be of adequate high quality for profitable use in reverse transcription PCR evaluation. Moreover, RNA isolates from feminine inflorescences had been used for the development of a cDNA library. This library was discovered to comprise a number of full-length cDNA clones of MADS-box genes, in line with the library being consultant of inflorescence expression profiles.

Transcriptome profiling of mouse samples utilizing nanopore sequencing of cDNA and RNA molecules.

Our imaginative and prescient of DNA transcription and splicing has modified dramatically with the introduction of short-read sequencing. These high-throughput sequencing applied sciences promised to unravel the complexity of any transcriptome. Typically gene expression ranges are well-captured utilizing these applied sciences, however there are nonetheless remaining caveats as a result of restricted learn size and the truth that RNA molecules needed to be reverse transcribed earlier than sequencing.

Oxford Nanopore Applied sciences has lately launched a transportable sequencer which provides the opportunity of sequencing lengthy reads and most significantly RNA molecules. Right here we generated a full mouse transcriptome from mind and liver utilizing the Oxford Nanopore machine. As a comparability, we sequenced RNA (RNA-Seq) and cDNA (cDNA-Seq) molecules utilizing each lengthy and quick reads applied sciences and examined the TeloPrime preparation package, devoted to the enrichment of full-length transcripts.

Utilizing spike-in knowledge, we confirmed that expression ranges are effectively captured by cDNA-Seq utilizing quick reads. Extra importantly, Oxford Nanopore RNA-Seq tends to be extra environment friendly, whereas cDNA-Seq seems to be extra biased. We additional present that the cDNA library preparation of the Nanopore protocol induces learn truncation for transcripts containing inside runs of T’s.

Extraction of high-quality tissue-specific RNA from London plane trees (Platanus acerifolia), permitting the construction of a female inflorescence cDNA library

This bias is marked for runs of at the very least 15 T’s, however is already detectable for runs of at the very least 9 T’s and due to this fact considerations greater than 20% of expressed transcripts in mouse mind and liver. Lastly, we define that bioinformatics challenges stay forward for quantifying on the transcript stage, particularly when reads are usually not full-length. Correct quantification of repeat-associated genes equivalent to processed pseudogenes additionally stays troublesome, and we present that present mapping protocols which map reads to the genome largely over-estimate their expression, on the expense of their mother or father gene.

Rolling Circle cDNA Synthesis Uncovers Round RNA Splice Variants.

Excessive-throughput RNA sequencing and novel bioinformatic pipelines have recognized 1000’s of round (circ)RNAs containing backsplice junction sequences. Nevertheless, circRNAs generated from a number of exons might comprise totally different combos of exons and/or introns arising from different splicing, whereas the backsplice junction sequence is similar. To have the ability to establish circRNA splice variants, we developed a way termed circRNA-Rolling Circle Amplification (circRNA-RCA).
This methodology detects full-length circRNA sequences by performing reverse transcription (RT) within the absence of RNase H exercise, adopted by polymerase chain response (PCR) amplification of full-length circRNAs utilizing a ahead primer spanning the backsplice junction sequence and a reverse primer precisely upstream of the ahead primer.
By sequencing the PCR merchandise, circRNA splice variants bearing the identical backsplice junctions, which had been in any other case solely predicted computationally, may very well be experimentally validated. The splice variants had been additional predicted to affiliate with totally different subsets of goal RNA-binding proteins and microRNAs, supporting the notion that totally different circRNA splice variants can have totally different organic impacts. In sum, the circRNA-RCA methodology permits the correct identification of full-length circRNA sequences, providing distinctive perception into their particular person operate.

Methods for Changing RNA to Amplifiable cDNA for Single-Cell RNA Sequencing Strategies.

This overview describes the options of molecular biology methods for single-cell RNA sequencing (scRNA-seq), together with strategies developed in our laboratory. Current scRNA-seq strategies require the conversion of first-strand cDNA to amplifiable cDNA adopted by whole-transcript amplification. There are three major methods for this conversion: poly-A tagging, template switching, and RNase H-DNA polymerase I-mediated second-strand cDNA synthesis for in vitro transcription.
We focus on the deserves and limitations of those methods and describe our Reverse Transcription with Random Displacement Amplification know-how that permits for direct first-strand cDNA amplification from RNA with out the necessity for conversion to an amplifiable cDNA.

RNASE9 cDNA Clone
MBS1267951-001mgPlasmid02mLGlycerolStock 0.01mgPlasmid+0.2mLGlycerol-Stock
EUR 200

RNASE9 cDNA Clone
MBS1267951-5x001mgPlasmid5x02mLGlycerolStock 5x0.01mgPlasmid+5x0.2mLGlycerol-Stock
EUR 855

RNASE1 cDNA Clone
MBS1269123-001mgPlasmid02mLGlycerolStock 0.01mgPlasmid+0.2mLGlycerol-Stock
EUR 200

RNASE1 cDNA Clone
MBS1269123-5x001mgPlasmid5x02mLGlycerolStock 5x0.01mgPlasmid+5x0.2mLGlycerol-Stock
EUR 855

RNASE4 cDNA Clone
MBS1276349-001mgPlasmid02mLGlycerolStock 0.01mgPlasmid+0.2mLGlycerol-Stock
EUR 200

RNASE4 cDNA Clone
MBS1276349-5x001mgPlasmid5x02mLGlycerolStock 5x0.01mgPlasmid+5x0.2mLGlycerol-Stock
EUR 855

RNASEL cDNA Clone
MBS1272159-001mgPlasmid02mLGlycerolStock 0.01mgPlasmid+0.2mLGlycerol-Stock
EUR 435

RNASEL cDNA Clone
MBS1272159-5x001mgPlasmid5x02mLGlycerolStock 5x0.01mgPlasmid+5x0.2mLGlycerol-Stock
EUR 1910

RNASET2 cDNA Clone
MBS1275311-001mgPlasmid02mLGlycerolStock 0.01mgPlasmid+0.2mLGlycerol-Stock
EUR 200

RNASET2 cDNA Clone
MBS1275311-5x001mgPlasmid5x02mLGlycerolStock 5x0.01mgPlasmid+5x0.2mLGlycerol-Stock
EUR 855

RNASEH2A cDNA Clone
MBS1272173-001mgPlasmid02mLGlycerolStock 0.01mgPlasmid+0.2mLGlycerol-Stock
EUR 315

RNASEH2A cDNA Clone
MBS1272173-5x001mgPlasmid5x02mLGlycerolStock 5x0.01mgPlasmid+5x0.2mLGlycerol-Stock
EUR 1385

RNASEH2B cDNA Clone
MBS1273020-001mgPlasmid02mLGlycerolStock 0.01mgPlasmid+0.2mLGlycerol-Stock
EUR 200

RNASEH2B cDNA Clone
MBS1273020-5x001mgPlasmid5x02mLGlycerolStock 5x0.01mgPlasmid+5x0.2mLGlycerol-Stock
EUR 855

RNASEH2C cDNA Clone
MBS1276698-001mgPlasmid02mLGlycerolStock 0.01mgPlasmid+0.2mLGlycerol-Stock
EUR 210

RNASEH2C cDNA Clone
MBS1276698-5x001mgPlasmid5x02mLGlycerolStock 5x0.01mgPlasmid+5x0.2mLGlycerol-Stock
EUR 905

Recombinant Ribonuclease A12 (RNASE12)
RPT642Hu01 10ug
EUR 174

Recombinant Ribonuclease A12 (RNASE12)
4-RPT642Hu01
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  • 5 mg
  • 10ug
  • 50ug
  • 100 ug
  • 200 ug
  • 500 ug
  • 1 mg
Description: Recombinant Human Ribonuclease A12 expressed in: E.coli

Polyclonal Antibody to Ribonuclease A12 (RNASE12)
PAT642Hu01 100ul
EUR 270

Ribonuclease A12 (RNASE12) Polyclonal Antibody (Human)
4-PAT642Hu01
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  • 20ul
  • 100ul
  • 200ul
  • 1ml
  • 10ml
Description: A Rabbit polyclonal antibody against Human Ribonuclease A12 (RNASE12)

Ribonuclease A12 (RNASE12) Polyclonal Antibody (Human), PE
4-PAT642Hu01-PE
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  • 20ul
  • 100ul
  • 200ul
  • 1ml
  • 10ml
Description: A Rabbit polyclonal antibody against Human Ribonuclease A12 (RNASE12). This antibody is labeled with PE.

Ribonuclease A12 (RNASE12) Polyclonal Antibody (Human), APC
4-PAT642Hu01-APC
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  • 20ul
  • 100ul
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  • 1ml
  • 10ml
Description: A Rabbit polyclonal antibody against Human Ribonuclease A12 (RNASE12). This antibody is labeled with APC.

Ribonuclease A12 (RNASE12) Polyclonal Antibody (Human), Cy3
4-PAT642Hu01-Cy3
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  • 20ul
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  • 1ml
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Description: A Rabbit polyclonal antibody against Human Ribonuclease A12 (RNASE12). This antibody is labeled with Cy3.

Ribonuclease A12 (RNASE12) Polyclonal Antibody (Human), HRP
4-PAT642Hu01-HRP
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  • 20ul
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  • 200ul
  • 1ml
  • 10ml
Description: A Rabbit polyclonal antibody against Human Ribonuclease A12 (RNASE12). This antibody is labeled with HRP.

Ribonuclease A12 (RNASE12) Polyclonal Antibody (Human), FITC
4-PAT642Hu01-FITC
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  • 20ul
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  • 1ml
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Description: A Rabbit polyclonal antibody against Human Ribonuclease A12 (RNASE12). This antibody is labeled with FITC.

Lenti ORF clone of Rnase12 (mGFP-tagged) - Mouse ribonuclease, RNase A family, 12 (non-active) (Rnase12)
MR215728L4 10 µg Ask for price

Ribonuclease A12 (RNASE12) Polyclonal Antibody (Human), Biotinylated
4-PAT642Hu01-Biotin
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Description: A Rabbit polyclonal antibody against Human Ribonuclease A12 (RNASE12). This antibody is labeled with Biotin.

Ribonuclease A12 (RNASE12) Polyclonal Antibody (Human), APC-Cy7
4-PAT642Hu01-APC-Cy7
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  • 20ul
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  • 1ml
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Description: A Rabbit polyclonal antibody against Human Ribonuclease A12 (RNASE12). This antibody is labeled with APC-Cy7.

Lenti ORF clone of Rnase12 (Myc-DDK-tagged) - Mouse ribonuclease, RNase A family, 12 (non-active) (Rnase12)
MR215728L3 10 µg Ask for price

Lenti ORF clone of Rnase12 (mGFP-tagged ORF) - Rat ribonuclease, RNase A family, 12 (non-active) (Rnase12), (10 ug)
RR204507L4 10 µg Ask for price

Lenti ORF clone of Rnase12 (Myc-DDK-tagged ORF) - Rat ribonuclease, RNase A family, 12 (non-active) (Rnase12), (10 ug)
RR204507L3 10 µg Ask for price

RNASE12 cloning plasmid
CSB-CL708031HU-10ug 10ug
EUR 280.8
Description: A cloning plasmid for the RNASE12 gene.

Rnase12/ Rat Rnase12 ELISA Kit
ELI-52551r 96 Tests
EUR 1063.2

RNASE12
CSB-CL708031HU 10 μg plasmid + 200μl Glycerol Ask for price

RNASE12
MBS8562224-01mLAF405L 0.1mL(AF405L)
EUR 565

RNASE12
MBS8562224-01mLAF405S 0.1mL(AF405S)
EUR 565

RNASE12
MBS8562224-01mLAF610 0.1mL(AF610)
EUR 565

RNASE12
MBS8562224-01mLAF635 0.1mL(AF635)
EUR 565

RNASE12
MBS8562224-02mL 0.2mL
EUR 345

RNASE11 (untagged)-Human ribonuclease, RNase A family, 11 (non-active) (cDNA clone MGC:33619 IMAGE:4826406), complete cds
SC122453 10 µg Ask for price

Lenti ORF clone of Human ribonuclease, RNase A family, 12 (non-active) (RNASE12), mGFP tagged
RC222112L4 10 µg Ask for price

RNASE12 antibody
38175 100ul
EUR 439

RNASE12 antibody
38175-100ul 100ul
EUR 302.4

RNASE12 Antibody
ABD6217 100 ug
EUR 525.6

RNASE12 Antibody
DF6217 200ul
EUR 420

RNASE12 Antibody
DF6217-100ul 100ul
EUR 168
Description: WB,IHC,IF/ICC,ELISA(peptide)

RNASE12 Antibody
DF6217-200ul 200ul
EUR 210
Description: WB,IHC,IF/ICC,ELISA(peptide)

RNASE12 Antibody
CSB-PA019792KA01HU- each
EUR 402
Description: A polyclonal antibody against RNASE12. Recognizes RNASE12 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, WB, IHC;WB:1:500-1:2000, IHC:1:50-1:200

RNASE12 Antibody
CSB-PA019792KA01HU-100ul 100ul
EUR 466.8
Description: A polyclonal antibody against RNASE12. Recognizes RNASE12 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, WB, IHC;WB:1:500-1:2000, IHC:1:50-1:200

RNASE12 Antibody
E309933 200ul
EUR 275
Description: Available in various conjugation types.

RNASE12 Antibody
E19-6217 100μg/100μl
EUR 225
Description: Available in various conjugation types.

RNASE12 Antibody
E91072 100ul
EUR 255
Description: Available in various conjugation types.

RNASE12 Antibody
MBS8501763-01mL 0.1mL
EUR 325

RNASE12 Antibody
MBS8501763-01mLAF405L 0.1mL(AF405L)
EUR 565

RNASE12 Antibody
MBS8501763-01mLAF405S 0.1mL(AF405S)
EUR 565

RNASE12 Antibody
MBS8501763-01mLAF610 0.1mL(AF610)
EUR 565

RNASE12 Antibody
MBS8501763-01mLAF635 0.1mL(AF635)
EUR 565

RNASE12 Antibody
MBS9606876-01mL 0.1mL
EUR 260

RNASE12 Antibody
MBS9606876-02mL 0.2mL
EUR 305

RNASE12 Antibody
MBS9606876-5x02mL 5x0.2mL
EUR 1220

RNASE12 Antibody
MBS8516134-01mg 0.1mg
EUR 305

RNASE12 Antibody
MBS8516134-01mLAF405L 0.1mL(AF405L)
EUR 465

RNASE12 Antibody
MBS8516134-01mLAF405S 0.1mL(AF405S)
EUR 465

RNASE12 Antibody
MBS8516134-01mLAF610 0.1mL(AF610)
EUR 465

RNASE12 Antibody
MBS8516134-01mLAF635 0.1mL(AF635)
EUR 465

RNASE12 antibody
MBS9401252-005mL 0.05mL
EUR 300

RNASE12 antibody
MBS9401252-01mL 0.1mL
EUR 390

RNASE12 antibody
MBS9401252-5x01mL 5x0.1mL
EUR 1610

Lenti ORF clone of Human ribonuclease, RNase A family, 12 (non-active) (RNASE12), Myc-DDK-tagged
RC222112L3 10 µg Ask for price

Rnaseh1 (untagged) - Mouse ribonuclease H1 (cDNA clone MGC:30220 IMAGE:5133942), (10ug)
MC206312 10 µg Ask for price

Rat RNASE12 siRNA
20-abx931614
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  • 15 nmol
  • 30 nmol

RNASE12 siRNA (Rat)
MBS8205845-15nmol 15nmol
EUR 405

RNASE12 siRNA (Rat)
MBS8205845-30nmol 30nmol
EUR 565

RNASE12 siRNA (Rat)
MBS8205845-5x30nmol 5x30nmol
EUR 2450

RNASE12 Rabbit pAb
A14607 50μL
EUR 262.15

RNASE12 Rabbit pAb
A14607-100ul 100 ul
EUR 369.6

RNASE12 Rabbit pAb
A14607-200ul 200 ul
EUR 550.8

RNASE12 Rabbit pAb
A14607-20ul 20 ul
EUR 219.6

RNASE12 Rabbit pAb
A14607-50ul 50 ul
EUR 267.6

RNASE12 Rabbit pAb
A14608 1000μL
EUR 262.15

RNASE12 Rabbit pAb
A14608-100ul 100 ul
EUR 369.6

RNASE12 Rabbit pAb
A14608-200ul 200 ul
EUR 550.8

RNASE12 Rabbit pAb
A14608-20ul 20 ul
EUR 219.6

RNASE12 Rabbit pAb
A14608-50ul 50 ul
EUR 267.6

RNASE12 Rabbit pAb
A1072 100μL
EUR 262.15

RNASE12 Rabbit pAb
A1072-100ul 100 ul
EUR 369.6

RNASE12 Rabbit pAb
A1072-200ul 200 ul
EUR 550.8

RNASE12 Rabbit pAb
A1072-20ul 20 ul
EUR 219.6

RNASE12 Rabbit pAb
A1072-50ul 50 ul
EUR 267.6

RNASE12 Rabbit pAb
E45R21165N 50 ul
EUR 297.75

RNASE12 Rabbit pAb
E45R21166N 50 ul
EUR 297.75

Human RNASE12 siRNA
20-abx904594
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  • 15 nmol
  • 30 nmol

Mouse RNASE12 siRNA
20-abx931615
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  • 15 nmol
  • 30 nmol

RNASE12 siRNA (Human)
MBS8206506-15nmol 15nmol
EUR 405

RNASE12 siRNA (Human)
MBS8206506-30nmol 30nmol
EUR 565

RNASE12 siRNA (Human)
MBS8206506-5x30nmol 5x30nmol
EUR 2450

RNASE12 siRNA (Mouse)
MBS8240121-15nmol 15nmol
EUR 405

RNASE12 siRNA (Mouse)
MBS8240121-30nmol 30nmol
EUR 565

RNASE12 siRNA (Mouse)
MBS8240121-5x30nmol 5x30nmol
EUR 2450

RNASE12 Blocking Peptide
DF6217-BP 1mg
EUR 234

RNASE12 Blocking Peptide
MBS9625593-1mg 1mg
EUR 380

RNASE12 Blocking Peptide
MBS9625593-5x1mg 5x1mg
EUR 1650

RNASE12 Polyclonal Antibody
E-AB-90905-120uL 120uL
EUR 320
Description: Unconjugated

RNASE12 Polyclonal Antibody
E-AB-90905-200uL 200uL
EUR 530
Description: Unconjugated

RNASE12 Polyclonal Antibody
E-AB-90905-60uL 60uL
EUR 200
Description: Unconjugated

RNASE12 Polyclonal Antibody
E-AB-90905-each each Ask for price
Description: Unconjugated

RNASE12 Polyclonal Antibody
E-AB-90906-120uL 120uL
EUR 320
Description: Unconjugated

RNASE12 Polyclonal Antibody
E-AB-90906-200uL 200uL
EUR 530
Description: Unconjugated

RNASE12 Polyclonal Antibody
E-AB-90906-60uL 60uL
EUR 200
Description: Unconjugated

RNASE12 Polyclonal Antibody
E-AB-90906-each each Ask for price
Description: Unconjugated

RNASE12 Conjugated Antibody
C38175 100ul
EUR 476.4

RNASE12 polyclonal antibody
BS75867 50ul
EUR 358
Description: 1mg/ml in PBS with 0.02% sodium azide, 50% glycerol, pH7.2

RNASE12 polyclonal antibody
BS75868 50ul
EUR 358
Description: 1mg/ml in PBS with 0.02% sodium azide, 50% glycerol, pH7.2

RNASE12 Polyclonal Antibody
E914607 100ul
EUR 225
Description: Available in various conjugation types.

RNASE12 Polyclonal Antibody
E914608 100ul
EUR 225
Description: Available in various conjugation types.

RNASE12 Polyclonal Antibody
MBS9134608-002mL 0.02mL
EUR 200

RNASE12 Polyclonal Antibody
MBS9134608-005mL 0.05mL
EUR 255

RNASE12 Polyclonal Antibody
MBS9134608-01mL 0.1mL
EUR 345

RNASE12 Polyclonal Antibody
MBS9134608-02mL 0.2mL
EUR 545

RNASE12 Polyclonal Antibody
MBS9134608-5x02mL 5x0.2mL
EUR 2265

RNASE12 Polyclonal Antibody
MBS9134609-002mL 0.02mL
EUR 200

RNASE12 Polyclonal Antibody
MBS9134609-005mL 0.05mL
EUR 255

RNASE12 Polyclonal Antibody
MBS9134609-01mL 0.1mL
EUR 345

RNASE12 Polyclonal Antibody
MBS9134609-02mL 0.2mL
EUR 545

RNASE12 Polyclonal Antibody
MBS9134609-5x02mL 5x0.2mL
EUR 2265

RNASE12 Polyclonal Antibody
MBS8569301-01mL 0.1mL
EUR 305
We imagine that this overview offers all customers of single-cell transcriptome applied sciences with an understanding of the connection between the quantitative efficiency of varied strategies and their molecular options.

 

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