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.

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.

RNASE12 Antibody
CSB-PA019792KA01HU-	Cusabio	EUR 335

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	Cusabio	100ul	EUR 389

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
DF6217	Affbiotech	200ul	EUR 304
Description: RNASE12 Antibody detects endogenous levels of total RNASE12.

RNASE12 Antibody
ABD6217	Lifescience Market	100 ug	EUR 438

RNASE12 Rabbit pAb
A1072-100ul	Abclonal	100 ul	EUR 308

RNASE12 Rabbit pAb
A1072-200ul	Abclonal	200 ul	EUR 459

RNASE12 Rabbit pAb
A1072-20ul	Abclonal	20 ul	EUR 183

RNASE12 Rabbit pAb
A1072-50ul	Abclonal	50 ul	EUR 223

RNASE12 Rabbit pAb
A14607-100ul	Abclonal	100 ul	EUR 308

RNASE12 Rabbit pAb
A14607-200ul	Abclonal	200 ul	EUR 459

RNASE12 Rabbit pAb
A14607-20ul	Abclonal	20 ul	EUR 183

RNASE12 Rabbit pAb
A14607-50ul	Abclonal	50 ul	EUR 223

RNASE12 Rabbit pAb
A14608-100ul	Abclonal	100 ul	EUR 308

RNASE12 Rabbit pAb
A14608-200ul	Abclonal	200 ul	EUR 459

RNASE12 Rabbit pAb
A14608-20ul	Abclonal	20 ul	EUR 183

RNASE12 Rabbit pAb
A14608-50ul	Abclonal	50 ul	EUR 223

Anti-RNASE12 antibody
STJ25363	St John's Laboratory	100 µl	EUR 277

Anti-RNASE12 antibody
STJ116817	St John's Laboratory	100 µl	EUR 277

Anti-RNASE12 antibody
STJ116818	St John's Laboratory	100 µl	EUR 277

RNASE12 cloning plasmid
CSB-CL708031HU-10ug	Cusabio	10ug	EUR 234

Description: A cloning plasmid for the RNASE12 gene.

RNASE12 Conjugated Antibody
C38175	SAB	100ul	EUR 397

RNASE12 Blocking Peptide
DF6217-BP	Affbiotech	1mg	EUR 195

Ribonuclease A12 (RNASE12) Polyclonal Antibody (Human)
4-PAT642Hu01	Cloud-Clone	EUR 262.00 EUR 2747.00 EUR 679.00 EUR 331.00 EUR 220.00	100ul 10ml 1ml 200ul 20ul

Description: A Rabbit polyclonal antibody against Human Ribonuclease A12 (RNASE12)

Ribonuclease A12 (RNASE12) Polyclonal Antibody (Human), APC
4-PAT642Hu01-APC	Cloud-Clone	EUR 368.00 EUR 3599.00 EUR 993.00 EUR 472.00 EUR 229.00	100ul 10ml 1ml 200ul 20ul

Description: A Rabbit polyclonal antibody against Human Ribonuclease A12 (RNASE12). This antibody is labeled with APC.

Ribonuclease A12 (RNASE12) Polyclonal Antibody (Human), Biotinylated
4-PAT642Hu01-Biotin	Cloud-Clone	EUR 328.00 EUR 2697.00 EUR 786.00 EUR 404.00 EUR 226.00	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), Cy3
4-PAT642Hu01-Cy3	Cloud-Clone	EUR 449.00 EUR 4757.00 EUR 1283.00 EUR 588.00 EUR 264.00	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), FITC
4-PAT642Hu01-FITC	Cloud-Clone	EUR 314.00 EUR 2899.00 EUR 814.00 EUR 397.00 EUR 203.00	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), HRP
4-PAT642Hu01-HRP	Cloud-Clone	EUR 335.00 EUR 3135.00 EUR 877.00 EUR 426.00 EUR 215.00	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), PE
4-PAT642Hu01-PE	Cloud-Clone	EUR 314.00 EUR 2899.00 EUR 814.00 EUR 397.00 EUR 203.00	100ul 10ml 1ml 200ul 20ul

Description: A Rabbit polyclonal antibody against Human Ribonuclease A12 (RNASE12). This antibody is labeled with PE.

Mouse RNASE12 shRNA Plasmid
20-abx984097	Abbexa	EUR 801.00 EUR 1121.00	150 µg 300 µg

Rat RNASE12 shRNA Plasmid
20-abx990586	Abbexa	EUR 801.00 EUR 1121.00	150 µg 300 µg

Ribonuclease A12 (RNASE12) Antibody
abx117029-100ug	Abbexa	100 ug	EUR 467

Ribonuclease A12 (RNASE12) Antibody
20-abx101918	Abbexa	EUR 453.00 EUR 133.00 EUR 1316.00 EUR 620.00 EUR 342.00	100 ug 10 ug 1 mg 200 ug 50 ug

Ribonuclease A12 (RNASE12) Antibody
20-abx000995	Abbexa	EUR 411.00 EUR 592.00 EUR 182.00 EUR 314.00	100 ul 200 ul 20 ul 50 ul

Ribonuclease A12 (RNASE12) Antibody
abx146450-100ug	Abbexa	100 ug	EUR 391

Human RNASE12 shRNA Plasmid
20-abx968360	Abbexa	EUR 801.00 EUR 1121.00	150 µg 300 µg

RNASE12 Recombinant Protein (Mouse)
RP168362	ABM	100 ug	Ask for price

RNASE12 Recombinant Protein (Rat)
RP226220	ABM	100 ug	Ask for price

RNASE12 Recombinant Protein (Human)
RP042925	ABM	100 ug	Ask for price

Ribonuclease A12 (RNASE12) Polyclonal Antibody (Human), APC-Cy7
4-PAT642Hu01-APC-Cy7	Cloud-Clone	EUR 616.00 EUR 7078.00 EUR 1867.00 EUR 824.00 EUR 338.00	100ul 10ml 1ml 200ul 20ul

Description: A Rabbit polyclonal antibody against Human Ribonuclease A12 (RNASE12). This antibody is labeled with APC-Cy7.

Human Ribonuclease A12 (RNASE12) Protein
20-abx068911	Abbexa	EUR 690.00 EUR 286.00 EUR 2110.00 EUR 815.00 EUR 495.00	100 ug 10 ug 1 mg 200 ug 50 ug

Ribonuclease A12 (RNASE12) Antibody (Biotin)
20-abx271884	Abbexa	EUR 453.00 EUR 244.00 EUR 1316.00 EUR 620.00 EUR 342.00	100 ug 10 ug 1 mg 200 ug 50 ug

Rnase12 ORF Vector (Mouse) (pORF)
ORF056122	ABM	1.0 ug DNA	EUR 506

Rnase12 ORF Vector (Rat) (pORF)
ORF075408	ABM	1.0 ug DNA	EUR 506

RNASE12 ORF Vector (Human) (pORF)
ORF014309	ABM	1.0 ug DNA	EUR 354

cDNA Synthesis SuperMix
20-abx09801420ulSystems	Abbexa	EUR 565.00 EUR 481.00	100 rxns × 20 ul Systems 50 rxns × 20 ul Systems

Novo? cDNA Kit
M1165-100	Biovision		EUR 354

Novo? cDNA Kit
M1165-25	Biovision		EUR 267

Evo? cDNA Supermix
M1168-100	Biovision		EUR 381

Evo? cDNA Supermix
M1168-25	Biovision		EUR 267

Novo? cDNA Supermix
M1169-100	Biovision		EUR 441

Novo? cDNA Supermix
M1169-25	Biovision		EUR 289

Human Ribonuclease A12(RNASE12)ELISA Kit
QY-E03175	Qayee Biotechnology	96T	EUR 361

RNASE12 sgRNA CRISPR Lentivector set (Human)
K1832201	ABM	3 x 1.0 ug	EUR 339

Rnase12 sgRNA CRISPR Lentivector set (Rat)
K6296901	ABM	3 x 1.0 ug	EUR 339

Rnase12 sgRNA CRISPR Lentivector set (Mouse)
K3570801	ABM	3 x 1.0 ug	EUR 339

cDNA from Plant Normal Tissue: cDNA from Plant: Arabidopsis
C1634310	Biochain	40 reactions	EUR 621
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

cDNA from Plant Normal Tissue: cDNA from Plant: Corn
C1634330	Biochain	40 reactions	EUR 621
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

cDNA from Plant Normal Tissue: cDNA from Plant: Orange
C1634340	Biochain	40 reactions	EUR 621
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

cDNA from Plant Normal Tissue: cDNA from Plant: Potato
C1634350	Biochain	40 reactions	EUR 621
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

cDNA from Plant Normal Tissue: cDNA from Plant: Rice
C1634360	Biochain	40 reactions	EUR 621
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

cDNA from Plant Normal Tissue: cDNA from Plant: Wheat
C1634390	Biochain	40 reactions	EUR 621
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

First-Strand cDNA Synthesis SuperMix (cDNA up to 12 kb)
20-abx09801620ulSystems	Abbexa	EUR 620.00 EUR 523.00	100 rxns × 20 ul Systems 50 rxns × 20 ul Systems

First-Strand cDNA Synthesis SuperMix (cDNA up to 15 kb)
20-abx09802120ulSystems	Abbexa	EUR 871.00 EUR 662.00	100 rxns × 20 ul Systems 50 rxns × 20 ul Systems

cDNA from Plant Normal Tissue: cDNA from Plant: Soy bean
C1634370	Biochain	40 reactions	EUR 621
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

cDNA Probe Diluent Solution
AR0063	BosterBio	5mL	EUR 106

Tetro cDNA Synthesis Kit
BIO-65042	Bioline	30 Reactions	Ask for price

Tetro cDNA Synthesis Kit
BIO-65043	Bioline	100 Reactions	Ask for price

SensiFAST cDNA Synthesis Kit
BIO-65053	Bioline	50 Reactions	Ask for price

SensiFAST cDNA Synthesis Kit
BIO-65053/S	Bioline	Sample	Ask for price

SensiFAST cDNA Synthesis Kit
BIO-65054	Bioline	250 Reactions	Ask for price

Human eNOS cDNA probe
eNOS51-D-2	Alpha Diagnostics	2 ug	EUR 445

Plant Tissue cDNA: Arabidopsis
PC34-310	Alpha Diagnostics	10 rxn	EUR 415

OneScriptPlus cDNA Synthesis Kit
G235	ABM	25 x 20 ul reactions	EUR 97

OneScriptPlus cDNA Synthesis Kit
G236	ABM	100 x 20 ul reactions	EUR 169

OneScriptPlus cDNA Synthesis SuperMix
G453	ABM	25 x 20 ul reactions	EUR 97

OneScriptPlus cDNA Synthesis SuperMix
G454	ABM	100 x 20 ul reactions	EUR 169

circRNA cDNA Synthesis Kit
G627	ABM	25 rxn (20 ul/rxn)	EUR 309

Evo™ cDNA Kit
M1164-100	Biovision		Ask for price

Evo™ cDNA Kit
M1164-25	Biovision		Ask for price

Novo? Transcriptome cDNA Kit
M1167-100	Biovision		EUR 952

Novo? Transcriptome cDNA Kit
M1167-25	Biovision		EUR 441

cDNA from Arteriosclerosis: Aorta
C1236012Hd-4	Biochain	40 reactions	EUR 811
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

cDNA from hypertension: Artery
C1236013Hd-2	Biochain	40 reactions	EUR 668
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

cDNA from Arteriosclerosis: Artery
C1236013Hd-4	Biochain	40 reactions	EUR 811
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

cDNA from hypertension: Vein
C1236020Hd-2	Biochain	40 reactions	EUR 668
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

cDNA from Lupus: Colon
C1236090Lup	Biochain	40 reactions	EUR 668
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

cDNA from hypertension: Heart
C1236122Hd-2	Biochain	40 reactions	EUR 668
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

cDNA from Lupus: Heart
C1236122Lup	Biochain	40 reactions	EUR 668
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

cDNA from hypertension: Kidney
C1236142Hd-2	Biochain	40 reactions	EUR 668
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

cDNA from Lupus: Kidney
C1236142Lup	Biochain	40 reactions	EUR 668
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

cDNA from Lupus: Liver
C1236149Lup	Biochain	40 reactions	EUR 668
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

cDNA from Asthma: Lung
C1236152Ld-1	Biochain	40 reactions	EUR 811
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

cDNA from Bronchitis: Lung
C1236152Ld-2	Biochain	40 reactions	EUR 811
Description: Can be used for various studies in the realm of gene expression, both normal and pathological. It is an excellent control and suitable for educational purposes.

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.