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

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.

Ribonuclease A12 (RNASE12) Polyclonal Antibody (Human), Cy3

4-PAT642Hu01-Cy3
  • EUR 538.80
  • EUR 5708.40
  • EUR 1539.60
  • EUR 705.60
  • EUR 316.80
  • 100ul
  • 10ml
  • 1ml
  • 200ul
  • 20ul
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
  • EUR 402.00
  • EUR 3762.00
  • EUR 1052.40
  • EUR 511.20
  • EUR 258.00
  • 100ul
  • 10ml
  • 1ml
  • 200ul
  • 20ul
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
  • EUR 376.80
  • EUR 3478.80
  • EUR 976.80
  • EUR 476.40
  • EUR 243.60
  • 100ul
  • 10ml
  • 1ml
  • 200ul
  • 20ul
Description: A Rabbit polyclonal antibody against Human Ribonuclease A12 (RNASE12). This antibody is labeled with FITC.

Ribonuclease A12 (RNASE12) Polyclonal Antibody (Human), Biotinylated

4-PAT642Hu01-Biotin
  • EUR 393.60
  • EUR 3236.40
  • EUR 943.20
  • EUR 484.80
  • EUR 271.20
  • 100ul
  • 10ml
  • 1ml
  • 200ul
  • 20ul
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
  • EUR 739.20
  • EUR 8493.60
  • EUR 2240.40
  • EUR 988.80
  • EUR 405.60
  • 100ul
  • 10ml
  • 1ml
  • 200ul
  • 20ul
Description: A Rabbit polyclonal antibody against Human Ribonuclease A12 (RNASE12). This antibody is labeled with APC-Cy7.

RNASE12 siRNA

20-abx904594
  • EUR 661.20
  • EUR 878.40
  • 15 nmol
  • 30 nmol

RNASE12 siRNA

20-abx931614
  • EUR 661.20
  • EUR 878.40
  • 15 nmol
  • 30 nmol

RNASE12 siRNA

20-abx931615
  • EUR 661.20
  • EUR 878.40
  • 15 nmol
  • 30 nmol

RNASE12 antibody

38175-100ul 100ul
EUR 302.4

RNASE12 Antibody

ABD6217 100 ug
EUR 525.6

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

DF6217-100ul 100ul
EUR 280

RNASE12 Antibody

DF6217-200ul 200ul
EUR 350

RNASE12 Antibody

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

RNASE12 Antibody

DF6217 100ul
EUR 280
Description: Human,Mouse,Rat

RNASE12 Antibody

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

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

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

A14608-100ul 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 Blocking Peptide

DF6217-BP 1mg
EUR 234

RNASE12 Conjugated Antibody

C38175 100ul
EUR 476.4

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.

Rat RNASE12 shRNA Plasmid

20-abx990586
  • EUR 961.20
  • EUR 1345.20
  • 150 µg
  • 300 µg

Mouse RNASE12 shRNA Plasmid

20-abx984097
  • EUR 961.20
  • EUR 1345.20
  • 150 µg
  • 300 µg

Human RNASE12 shRNA Plasmid

20-abx968360
  • EUR 961.20
  • EUR 1345.20
  • 150 µg
  • 300 µg

RNASE12 Recombinant Protein (Rat)

RP226220 100 ug Ask for price

Ribonuclease A12 (RNASE12) Antibody

abx117029-100ug 100 ug
EUR 560.4

Ribonuclease A12 (RNASE12) Antibody

abx146450-100ug 100 ug
EUR 469.2

Ribonuclease A12 (RNASE12) Antibody

20-abx101918
  • EUR 543.60
  • EUR 159.60
  • EUR 1579.20
  • EUR 744.00
  • EUR 410.40
  • 100 ug
  • 10 ug
  • 1 mg
  • 200 ug
  • 50 ug

Ribonuclease A12 (RNASE12) Antibody

20-abx000995
  • EUR 493.20
  • EUR 710.40
  • EUR 218.40
  • EUR 376.80
  • 100 ul
  • 200 ul
  • 20 ul
  • 50 ul

RNASE12 Recombinant Protein (Mouse)

RP168362 100 ug Ask for price

RNASE12 Recombinant Protein (Human)

RP042925 100 ug Ask for price

Recombinant Ribonuclease A12 (RNASE12)

RPU43641-100ug 100ug
EUR 537.9

Recombinant Ribonuclease A12 (RNASE12)

RPU43641-1mg 1mg
EUR 2380.3

Recombinant Ribonuclease A12 (RNASE12)

RPU43641-50ug 50ug
EUR 431.2

Rnase12 ORF Vector (Rat) (pORF)

ORF075408 1.0 ug DNA
EUR 607.2

RNASE12 ORF Vector (Human) (pORF)

ORF014309 1.0 ug DNA
EUR 424.8

Rnase12 ORF Vector (Mouse) (pORF)

ORF056122 1.0 ug DNA
EUR 607.2

Human RNASE12 Protein Lysate 20ug

IHURNASE12PLLY20UG each
EUR 213
Description: Human RNASE12 Protein Lysate 20ug

Human Ribonuclease A12 (RNASE12) Protein

20-abx068911
  • EUR 828.00
  • EUR 343.20
  • EUR 2532.00
  • EUR 978.00
  • EUR 594.00
  • 100 ug
  • 10 ug
  • 1 mg
  • 200 ug
  • 50 ug

Ribonuclease A12 (RNASE12) Antibody (Biotin)

20-abx271884
  • EUR 543.60
  • EUR 292.80
  • EUR 1579.20
  • EUR 744.00
  • EUR 410.40
  • 100 ug
  • 10 ug
  • 1 mg
  • 200 ug
  • 50 ug

Genorise® Chicken TNFa cDNA Clone

GR147001 - Ask for price

Genorise® Chicken IFNg cDNA Clone

GR147002 10 µg
EUR 153

Ribonuclease A12 (RNASE12) Polyclonal Antibody

CAU21029-100ul 100ul
EUR 259.2
Description: polyclonal

Ribonuclease A12 (RNASE12) Polyclonal Antibody

CAU21029-200ul 200ul
EUR 324.2
Description: polyclonal

Rnase12 sgRNA CRISPR Lentivector set (Rat)

K6296901 3 x 1.0 ug
EUR 406.8

Human Ribonuclease A12(RNASE12)ELISA Kit

QY-E03175 96T
EUR 433.2

Rnase12 3'UTR GFP Stable Cell Line

TU269478 1.0 ml Ask for price

Rnase12 3'UTR GFP Stable Cell Line

TU167925 1.0 ml Ask for price

RNASE12 3'UTR GFP Stable Cell Line

TU070002 1.0 ml
EUR 2799.6

RNASE12 sgRNA CRISPR Lentivector set (Human)

K1832201 3 x 1.0 ug
EUR 406.8

Rnase12 sgRNA CRISPR Lentivector set (Mouse)

K3570801 3 x 1.0 ug
EUR 406.8

Genorise® Chicken IL-6 cDNA Clone

GR147003 10 µg
EUR 153

Genorise® Chicken IL-4 cDNA Clone

GR147004 10 µg
EUR 153

Genorise® Chicken IL-10 cDNA Clone

GR147005 10 µg
EUR 153

RNASE12 Protein Vector (Rat) (pPM-C-HA)

PV301632 500 ng
EUR 723.6

RNASE12 Protein Vector (Rat) (pPB-C-His)

PV301630 500 ng
EUR 723.6

RNASE12 Protein Vector (Rat) (pPB-N-His)

PV301631 500 ng
EUR 723.6

RNASE12 Protein Vector (Rat) (pPM-C-His)

PV301633 500 ng
EUR 723.6

RNASE12 Protein Vector (Human) (pPM-C-HA)

PV057235 500 ng
EUR 577.2

RNASE12 Protein Vector (Mouse) (pPM-C-HA)

PV224488 500 ng
EUR 723.6

RNASE12 Protein Vector (Human) (pPB-C-His)

PV057233 500 ng
EUR 577.2

RNASE12 Protein Vector (Human) (pPB-N-His)

PV057234 500 ng
EUR 577.2

RNASE12 Protein Vector (Human) (pPM-C-His)

PV057236 500 ng
EUR 577.2

RNASE12 Protein Vector (Mouse) (pPB-C-His)

PV224486 500 ng
EUR 723.6

RNASE12 Protein Vector (Mouse) (pPB-N-His)

PV224487 500 ng
EUR 723.6

RNASE12 Protein Vector (Mouse) (pPM-C-His)

PV224489 500 ng
EUR 723.6

Rnase12 3'UTR Luciferase Stable Cell Line

TU219478 1.0 ml Ask for price

Rnase12 3'UTR Luciferase Stable Cell Line

TU117925 1.0 ml Ask for price

RNASE12 3'UTR Luciferase Stable Cell Line

TU020002 1.0 ml
EUR 2799.6

Rnase12 sgRNA CRISPR Lentivector (Rat) (Target 1)

K6296902 1.0 ug DNA
EUR 184.8

Rnase12 sgRNA CRISPR Lentivector (Rat) (Target 2)

K6296903 1.0 ug DNA
EUR 184.8

Rnase12 sgRNA CRISPR Lentivector (Rat) (Target 3)

K6296904 1.0 ug DNA
EUR 184.8

Genorise® Human BNP cDNA Clone, 10 ug

GR154005 10 µg
EUR 153

Genorise® Human ANP cDNA Clone, 10 ug

GR154008 10 µg
EUR 153

Genorise® Human CRP cDNA Clone, 10 ug

GR154011 10 µg
EUR 153

Genorise® Human cTnT cDNA Clone, 10 ug

GR154003 10 µg
EUR 153

Genorise® Human TNN3 cDNA Clone, 10 ug

GR154004 10 µg
EUR 153

Genorise® Human GPBB cDNA Clone, 10 ug

GR154010 10 µg
EUR 153

RNASE12 sgRNA CRISPR Lentivector (Human) (Target 1)

K1832202 1.0 ug DNA
EUR 184.8

RNASE12 sgRNA CRISPR Lentivector (Human) (Target 2)

K1832203 1.0 ug DNA
EUR 184.8

RNASE12 sgRNA CRISPR Lentivector (Human) (Target 3)

K1832204 1.0 ug DNA
EUR 184.8

Rnase12 sgRNA CRISPR Lentivector (Mouse) (Target 1)

K3570802 1.0 ug DNA
EUR 184.8

Rnase12 sgRNA CRISPR Lentivector (Mouse) (Target 2)

K3570803 1.0 ug DNA
EUR 184.8

Rnase12 sgRNA CRISPR Lentivector (Mouse) (Target 3)

K3570804 1.0 ug DNA
EUR 184.8

Genorise® Human GLUT1 cDNA Clone, 10 ug

GR154002 10 µg
EUR 153

Genorise® Human MyBPC3 cDNA Clone, 10 ug

GR154012 10 µg
EUR 153

Genorise® Human Myoglobin cDNA Clone, 10 ug

GR154006 10 µg
EUR 153

Genorise® Sheep IL-6 cDNA Clone, 10 ug

GR151001 10 µg
EUR 153

Genorise® Ovine IL-2 cDNA Clone, 10 ug

GR151002 10 µg
EUR 153

Genorise® Ovine IL-4 cDNA Clone, 10 ug

GR151003 10 µg
EUR 153

Mouse Ribonuclease- like protein 12, Rnase12 ELISA KIT

ELI-38874m 96 Tests
EUR 1038

Human Ribonuclease- like protein 12, RNASE12 ELISA KIT

ELI-18518h 96 Tests
EUR 988.8

Genorise® Canine IL-7 cDNA Clone, 10 ug

GR159002 10 µg
EUR 153

RNASE12 Lentiviral Vector (Rat) (CMV) (pLenti-GIII-CMV)

LV692671 1.0 ug DNA
EUR 616.8

RNASE12 Lentiviral Vector (Rat) (UbC) (pLenti-GIII-UbC)

LV692675 1.0 ug DNA
EUR 616.8

Genorise® Bovine IL-1b cDNA Clone, 10 ug

GR152001 10 µg
EUR 153

Genorise® Rabbit IL-1b cDNA Clone, 10 ug

GR153001 10 µg
EUR 153

Genorise® Human H-FABP cDNA Clone, 10 ug

GR154001 10 µg
EUR 153

Genorise® Human PAPP-A cDNA Clone, 10 ug

GR154013 10 µg
EUR 153

Genorise® Human Pro-C3 cDNA Clone, 10 ug

GR154015 10 µg
EUR 153

Genorise® Canine NGF-B cDNA Clone, 10 ug

GR159001 10 µg
EUR 153

RNASE12 Lentiviral Vector (Rat) (EF1a) (pLenti-GIII-EF1a)

LV692676 1.0 ug DNA
EUR 616.8

Genorise® Human Lp-PLA2 cDNA Clone, 10 ug

GR154007 10 µg
EUR 153

Genorise® Human Myeloperoxidase cDNA Clone, 10 ug

GR154009 10 µg
EUR 153

Genorise® Human NT-proBNP cDNA Clone, 10 ug

GR154014 10 µg
EUR 153

Rnase12 sgRNA CRISPR/Cas9 All-in-One Lentivector set (Rat)

K6296905 3 x 1.0 ug
EUR 451.2

RNASE12 sgRNA CRISPR/Cas9 All-in-One Lentivector set (Human)

K1832205 3 x 1.0 ug
EUR 451.2

Rnase12 sgRNA CRISPR/Cas9 All-in-One Lentivector set (Mouse)

K3570805 3 x 1.0 ug
EUR 451.2

Recombinant Ribonuclease A10 (RNASE10)

4-RPQ746Hu01
  • EUR 588.79
  • EUR 280.80
  • EUR 1877.95
  • EUR 705.98
  • EUR 1291.97
  • EUR 469.20
  • EUR 4514.88
  • 100 ug
  • 10ug
  • 1 mg
  • 200 ug
  • 500 ug
  • 50ug
  • 5 mg
Description: Recombinant Human Ribonuclease A10 expressed in: E.coli

Recombinant Ribonuclease A11 (RNASE11)

4-RPT488Hu01
  • EUR 588.79
  • EUR 280.80
  • EUR 1877.95
  • EUR 705.98
  • EUR 1291.97
  • EUR 469.20
  • EUR 4514.88
  • 100 ug
  • 10ug
  • 1 mg
  • 200 ug
  • 500 ug
  • 50ug
  • 5 mg
Description: Recombinant Human Ribonuclease A11 expressed in: E.coli

Recombinant Ribonuclease A13 (RNASE13)

4-RPT665Hu01
  • EUR 636.10
  • EUR 294.00
  • EUR 2055.36
  • EUR 765.12
  • EUR 1410.24
  • EUR 501.60
  • EUR 4958.40
  • 100 ug
  • 10ug
  • 1 mg
  • 200 ug
  • 500 ug
  • 50ug
  • 5 mg
Description: Recombinant Human Ribonuclease A13 expressed in: E.coli

Rnase12 sgRNA CRISPR/Cas9 All-in-One Lentivector (Rat) (Target 1)

K6296906 1.0 ug DNA
EUR 200.4

Rnase12 sgRNA CRISPR/Cas9 All-in-One Lentivector (Rat) (Target 2)

K6296907 1.0 ug DNA
EUR 200.4
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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