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.

Ribonuclease A12 (RNASE12) Polyclonal Antibody (Human), Cy3
4-PAT642Hu01-Cy3	Cloud-Clone	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	Cloud-Clone	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	Cloud-Clone	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	Cloud-Clone	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	Cloud-Clone	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	Abbexa	EUR 661.20 EUR 878.40	15 nmol 30 nmol

RNASE12 siRNA
20-abx931614	Abbexa	EUR 661.20 EUR 878.40	15 nmol 30 nmol

RNASE12 siRNA
20-abx931615	Abbexa	EUR 661.20 EUR 878.40	15 nmol 30 nmol

RNASE12 antibody
38175-100ul	SAB	100ul	EUR 302.4

RNASE12 Antibody
ABD6217	Lifescience Market	100 ug	EUR 525.6

RNASE12 Antibody
CSB-PA019792KA01HU-	Cusabio	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	Cusabio	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	EnoGene	200ul	EUR 275
Description: Available in various conjugation types.

RNASE12 Antibody
DF6217-100ul	Affinity Biosciences	100ul	EUR 280

RNASE12 Antibody
DF6217-200ul	Affinity Biosciences	200ul	EUR 350

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

RNASE12 Antibody
DF6217	Affinity Biosciences	100ul	EUR 280
Description: Human,Mouse,Rat

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

RNASE12 Rabbit pAb
A1072-100ul	Abclonal	100 ul	EUR 369.6

RNASE12 Rabbit pAb
A1072-200ul	Abclonal	200 ul	EUR 550.8

RNASE12 Rabbit pAb
A1072-20ul	Abclonal	20 ul	EUR 219.6

RNASE12 Rabbit pAb
A1072-50ul	Abclonal	50 ul	EUR 267.6

RNASE12 Rabbit pAb
A14607-100ul	Abclonal	100 ul	EUR 369.6

RNASE12 Rabbit pAb
A14607-200ul	Abclonal	200 ul	EUR 550.8

RNASE12 Rabbit pAb
A14607-20ul	Abclonal	20 ul	EUR 219.6

RNASE12 Rabbit pAb
A14607-50ul	Abclonal	50 ul	EUR 267.6

RNASE12 Rabbit pAb
A14608-100ul	Abclonal	100 ul	EUR 369.6

RNASE12 Rabbit pAb
A14608-200ul	Abclonal	200 ul	EUR 550.8

RNASE12 Rabbit pAb
A14608-20ul	Abclonal	20 ul	EUR 219.6

RNASE12 Rabbit pAb
A14608-50ul	Abclonal	50 ul	EUR 267.6

RNASE12 Blocking Peptide
DF6217-BP	Affbiotech	1mg	EUR 234

RNASE12 Conjugated Antibody
C38175	SAB	100ul	EUR 476.4

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

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

Rat RNASE12 shRNA Plasmid
20-abx990586	Abbexa	EUR 961.20 EUR 1345.20	150 µg 300 µg

Mouse RNASE12 shRNA Plasmid
20-abx984097	Abbexa	EUR 961.20 EUR 1345.20	150 µg 300 µg

Human RNASE12 shRNA Plasmid
20-abx968360	Abbexa	EUR 961.20 EUR 1345.20	150 µg 300 µg

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

Ribonuclease A12 (RNASE12) Antibody
abx117029-100ug	Abbexa	100 ug	EUR 560.4

Ribonuclease A12 (RNASE12) Antibody
abx146450-100ug	Abbexa	100 ug	EUR 469.2

Ribonuclease A12 (RNASE12) Antibody
20-abx101918	Abbexa	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	Abbexa	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	ABM	100 ug	Ask for price

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

Recombinant Ribonuclease A12 (RNASE12)
RPU43641-100ug	Biomatik Corporation	100ug	EUR 537.9

Recombinant Ribonuclease A12 (RNASE12)
RPU43641-1mg	Biomatik Corporation	1mg	EUR 2380.3

Recombinant Ribonuclease A12 (RNASE12)
RPU43641-50ug	Biomatik Corporation	50ug	EUR 431.2

Rnase12 ORF Vector (Rat) (pORF)
ORF075408	ABM	1.0 ug DNA	EUR 607.2

RNASE12 ORF Vector (Human) (pORF)
ORF014309	ABM	1.0 ug DNA	EUR 424.8

Rnase12 ORF Vector (Mouse) (pORF)
ORF056122	ABM	1.0 ug DNA	EUR 607.2

Human RNASE12 Protein Lysate 20ug
IHURNASE12PLLY20UG	Innovative research	each	EUR 213

Description: Human RNASE12 Protein Lysate 20ug

Human Ribonuclease A12 (RNASE12) Protein
20-abx068911	Abbexa	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	Abbexa	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	Genorise Scientific	-	Ask for price

Genorise® Chicken IFNg cDNA Clone
GR147002	Genorise Scientific	10 µg	EUR 153

Ribonuclease A12 (RNASE12) Polyclonal Antibody
CAU21029-100ul	Biomatik Corporation	100ul	EUR 259.2
Description: polyclonal

Ribonuclease A12 (RNASE12) Polyclonal Antibody
CAU21029-200ul	Biomatik Corporation	200ul	EUR 324.2
Description: polyclonal

Rnase12 sgRNA CRISPR Lentivector set (Rat)
K6296901	ABM	3 x 1.0 ug	EUR 406.8

Human Ribonuclease A12(RNASE12)ELISA Kit
QY-E03175	Qayee Biotechnology	96T	EUR 433.2

Rnase12 3'UTR GFP Stable Cell Line
TU269478	ABM	1.0 ml	Ask for price

Rnase12 3'UTR GFP Stable Cell Line
TU167925	ABM	1.0 ml	Ask for price

RNASE12 3'UTR GFP Stable Cell Line
TU070002	ABM	1.0 ml	EUR 2799.6

RNASE12 sgRNA CRISPR Lentivector set (Human)
K1832201	ABM	3 x 1.0 ug	EUR 406.8

Rnase12 sgRNA CRISPR Lentivector set (Mouse)
K3570801	ABM	3 x 1.0 ug	EUR 406.8

Genorise® Chicken IL-6 cDNA Clone
GR147003	Genorise Scientific	10 µg	EUR 153

Genorise® Chicken IL-4 cDNA Clone
GR147004	Genorise Scientific	10 µg	EUR 153

Genorise® Chicken IL-10 cDNA Clone
GR147005	Genorise Scientific	10 µg	EUR 153

RNASE12 Protein Vector (Rat) (pPM-C-HA)
PV301632	ABM	500 ng	EUR 723.6

RNASE12 Protein Vector (Rat) (pPB-C-His)
PV301630	ABM	500 ng	EUR 723.6

RNASE12 Protein Vector (Rat) (pPB-N-His)
PV301631	ABM	500 ng	EUR 723.6

RNASE12 Protein Vector (Rat) (pPM-C-His)
PV301633	ABM	500 ng	EUR 723.6

RNASE12 Protein Vector (Human) (pPM-C-HA)
PV057235	ABM	500 ng	EUR 577.2

RNASE12 Protein Vector (Mouse) (pPM-C-HA)
PV224488	ABM	500 ng	EUR 723.6

RNASE12 Protein Vector (Human) (pPB-C-His)
PV057233	ABM	500 ng	EUR 577.2

RNASE12 Protein Vector (Human) (pPB-N-His)
PV057234	ABM	500 ng	EUR 577.2

RNASE12 Protein Vector (Human) (pPM-C-His)
PV057236	ABM	500 ng	EUR 577.2

RNASE12 Protein Vector (Mouse) (pPB-C-His)
PV224486	ABM	500 ng	EUR 723.6

RNASE12 Protein Vector (Mouse) (pPB-N-His)
PV224487	ABM	500 ng	EUR 723.6

RNASE12 Protein Vector (Mouse) (pPM-C-His)
PV224489	ABM	500 ng	EUR 723.6

Rnase12 3'UTR Luciferase Stable Cell Line
TU219478	ABM	1.0 ml	Ask for price

Rnase12 3'UTR Luciferase Stable Cell Line
TU117925	ABM	1.0 ml	Ask for price

RNASE12 3'UTR Luciferase Stable Cell Line
TU020002	ABM	1.0 ml	EUR 2799.6

Rnase12 sgRNA CRISPR Lentivector (Rat) (Target 1)
K6296902	ABM	1.0 ug DNA	EUR 184.8

Rnase12 sgRNA CRISPR Lentivector (Rat) (Target 2)
K6296903	ABM	1.0 ug DNA	EUR 184.8

Rnase12 sgRNA CRISPR Lentivector (Rat) (Target 3)
K6296904	ABM	1.0 ug DNA	EUR 184.8

Genorise® Human BNP cDNA Clone, 10 ug
GR154005	Genorise Scientific	10 µg	EUR 153

Genorise® Human ANP cDNA Clone, 10 ug
GR154008	Genorise Scientific	10 µg	EUR 153

Genorise® Human CRP cDNA Clone, 10 ug
GR154011	Genorise Scientific	10 µg	EUR 153

Genorise® Human cTnT cDNA Clone, 10 ug
GR154003	Genorise Scientific	10 µg	EUR 153

Genorise® Human TNN3 cDNA Clone, 10 ug
GR154004	Genorise Scientific	10 µg	EUR 153

Genorise® Human GPBB cDNA Clone, 10 ug
GR154010	Genorise Scientific	10 µg	EUR 153

RNASE12 sgRNA CRISPR Lentivector (Human) (Target 1)
K1832202	ABM	1.0 ug DNA	EUR 184.8

RNASE12 sgRNA CRISPR Lentivector (Human) (Target 2)
K1832203	ABM	1.0 ug DNA	EUR 184.8

RNASE12 sgRNA CRISPR Lentivector (Human) (Target 3)
K1832204	ABM	1.0 ug DNA	EUR 184.8

Rnase12 sgRNA CRISPR Lentivector (Mouse) (Target 1)
K3570802	ABM	1.0 ug DNA	EUR 184.8

Rnase12 sgRNA CRISPR Lentivector (Mouse) (Target 2)
K3570803	ABM	1.0 ug DNA	EUR 184.8

Rnase12 sgRNA CRISPR Lentivector (Mouse) (Target 3)
K3570804	ABM	1.0 ug DNA	EUR 184.8

Genorise® Human GLUT1 cDNA Clone, 10 ug
GR154002	Genorise Scientific	10 µg	EUR 153

Genorise® Human MyBPC3 cDNA Clone, 10 ug
GR154012	Genorise Scientific	10 µg	EUR 153

Genorise® Human Myoglobin cDNA Clone, 10 ug
GR154006	Genorise Scientific	10 µg	EUR 153

Genorise® Sheep IL-6 cDNA Clone, 10 ug
GR151001	Genorise Scientific	10 µg	EUR 153

Genorise® Ovine IL-2 cDNA Clone, 10 ug
GR151002	Genorise Scientific	10 µg	EUR 153

Genorise® Ovine IL-4 cDNA Clone, 10 ug
GR151003	Genorise Scientific	10 µg	EUR 153

Mouse Ribonuclease- like protein 12, Rnase12 ELISA KIT
ELI-38874m	Lifescience Market	96 Tests	EUR 1038

Human Ribonuclease- like protein 12, RNASE12 ELISA KIT
ELI-18518h	Lifescience Market	96 Tests	EUR 988.8

Genorise® Canine IL-7 cDNA Clone, 10 ug
GR159002	Genorise Scientific	10 µg	EUR 153

RNASE12 Lentiviral Vector (Rat) (CMV) (pLenti-GIII-CMV)
LV692671	ABM	1.0 ug DNA	EUR 616.8

RNASE12 Lentiviral Vector (Rat) (UbC) (pLenti-GIII-UbC)
LV692675	ABM	1.0 ug DNA	EUR 616.8

Genorise® Bovine IL-1b cDNA Clone, 10 ug
GR152001	Genorise Scientific	10 µg	EUR 153

Genorise® Rabbit IL-1b cDNA Clone, 10 ug
GR153001	Genorise Scientific	10 µg	EUR 153

Genorise® Human H-FABP cDNA Clone, 10 ug
GR154001	Genorise Scientific	10 µg	EUR 153

Genorise® Human PAPP-A cDNA Clone, 10 ug
GR154013	Genorise Scientific	10 µg	EUR 153

Genorise® Human Pro-C3 cDNA Clone, 10 ug
GR154015	Genorise Scientific	10 µg	EUR 153

Genorise® Canine NGF-B cDNA Clone, 10 ug
GR159001	Genorise Scientific	10 µg	EUR 153

RNASE12 Lentiviral Vector (Rat) (EF1a) (pLenti-GIII-EF1a)
LV692676	ABM	1.0 ug DNA	EUR 616.8

Genorise® Human Lp-PLA2 cDNA Clone, 10 ug
GR154007	Genorise Scientific	10 µg	EUR 153

Genorise® Human Myeloperoxidase cDNA Clone, 10 ug
GR154009	Genorise Scientific	10 µg	EUR 153

Genorise® Human NT-proBNP cDNA Clone, 10 ug
GR154014	Genorise Scientific	10 µg	EUR 153

Rnase12 sgRNA CRISPR/Cas9 All-in-One Lentivector set (Rat)
K6296905	ABM	3 x 1.0 ug	EUR 451.2

RNASE12 sgRNA CRISPR/Cas9 All-in-One Lentivector set (Human)
K1832205	ABM	3 x 1.0 ug	EUR 451.2

Rnase12 sgRNA CRISPR/Cas9 All-in-One Lentivector set (Mouse)
K3570805	ABM	3 x 1.0 ug	EUR 451.2

Recombinant Ribonuclease A10 (RNASE10)
4-RPQ746Hu01	Cloud-Clone	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	Cloud-Clone	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	Cloud-Clone	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	ABM	1.0 ug DNA	EUR 200.4

Rnase12 sgRNA CRISPR/Cas9 All-in-One Lentivector (Rat) (Target 2)
K6296907	ABM	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.