A non-radioactive, improved PAR-CLIP and small RNA cDNA library preparation protocol

A non-radioactive, improved PAR-CLIP and small RNA cDNA library preparation protocol

Crosslinking and immunoprecipitation (CLIP) strategies are highly effective strategies to interrogate direct protein-RNA interactions and dissect posttranscriptional gene regulatory networks. One broadly used CLIP variant is photoactivatable ribonucleoside enhanced CLIP (PAR-CLIP) that entails in vivo labeling of nascent RNAs with the photoreactive nucleosides 4-thiouridine (4SU) or 6-thioguanosine (6SG), which might effectively crosslink to interacting proteins utilizing UVA and UVB mild.

Crosslinking of 4SU or 6SG to interacting amino acids adjustments their base-pairing properties and leads to attribute mutations in cDNA libraries ready for high-throughput sequencing, which could be computationally exploited to take away ample background from non-crosslinked sequences and assist pinpoint RNA binding protein binding websites at nucleotide decision on a transcriptome-wide scale.

Right here we current a streamlined protocol for fluorescence-based PAR-CLIP (fPAR-CLIP) that eliminates the necessity to use radioactivity. It’s primarily based on direct ligation of a fluorescently labeled adapter to the three’finish of crosslinked RNA on immobilized ribonucleoproteins, adopted by isolation of the adapter-ligated RNA and environment friendly conversion into cDNA with out the beforehand wanted measurement fractionation on denaturing polyacrylamide gels. These enhancements reduce the experimentation by half to 2 days and will increase sensitivity by 10-100-fold.

Characterization and evaluation of the transcriptome response to drought in Larix kaempferi utilizing PacBio full-length cDNA sequencing built-in with de novo RNA-seq reads

A hypothetical mannequin of drought tolerance mechanism of Larix kaempferi was established via SMRT-seq and Illumina HiSeq. Larix kaempferi is a vital financial and ecological species and a significant afforestation species in north-eastern China. Up to now, no info has been reliably derived concerning full-length cDNA sequencing info on L. kaempferi. By single-molecule long-read isoform sequencing (SMRT-seq), right here we report a complete of 26,153,342 subreads (21.24 Gb) and 330,371 round consensus sequence (CCS) reads after the modification of web site mismatch, and 35,414 unigenes have been efficiently collected.

To achieve deeper insights into the molecular mechanisms of L. kaempferi response to drought stress, we mixed Illumina HiSeq with SMRT-seq to decode full-length transcripts. On this examine, we report 27 differentially expressed genes (DEGs) concerned within the notion and transmission of drought stress indicators in L. kaempferi.

Numerous DEGs responding to drought stress have been detected in L. kaempferi, particularly DEGs concerned within the reactive oxygen species (ROS) scavenging, lignin biosynthesis, and sugar metabolism, and DEGs encoding drought stress proteins. We detected 73 transcription components (TFs) below drought stress, together with AP2/ERF, bZIP, TCP, and MYB. This examine offers primary full sequence sources for L. kaempferi analysis and can assist us to raised perceive the capabilities of drought-resistance genes in L. kaempferi.

A non-radioactive, improved PAR-CLIP and small RNA cDNA library preparation protocol

cDNA-derived RNA phage meeting reveals important residues within the maturation protein of the Pseudomonas aeruginosa leviphage, PP7

PP7 is a leviphage with single-stranded RNA genome, which infects Pseudomonas aeruginosa PAO1. A reverse genetic system for PP7 was beforehand created through the use of reverse-transcribed cDNA (PP7O) from virion-derived RNA genome. Right here, we’ve discovered that the PP7O cDNA contained 20 nucleotide variations from the PP7 genome sequence deposited within the database.

We created one other reverse genetic system exploiting chemically synthesized cDNA (PP7S) primarily based on the database sequence. In contrast to PP7O that rendered infectious PP7 virions, PP7S-derived particles have been incapable of plaque formation on PAO1 cells, which was restored on the PAO1 cells expressing the maturation protein (MP) from PP7O Utilizing this reverse genetic system, we revealed two amino acid residues concerned within the identified roles of MP (i.e. adsorption and genome replication), fortuitously offering a lesson that the viral RNA genome sequencing wants purposeful verification presumably by a reverse genetic system.

IMPORTANCE Organic significance of RNA phages has been largely ignored, mockingly as a result of few research have been specializing in RNA phages. As an preliminary try to correctly signify RNA phages within the phageome, we beforehand created, through the use of reverse-transcribed cDNA, a reverse genetic system for the small RNA phage, PP7 that infects the opportunistic human pathogen, Pseudomonas aeruginosa We right here report one other system through the use of chemically synthesized cDNA primarily based on the database genome that has 20 nucleotide variations from the earlier cDNA.

Investigation of these cDNA-derived phage virions unveiled that two amino acids of the maturation protein are essential for the traditional phage lifecycle at totally different steps. Our examine offers an perception into the molecular foundation for the RNA phage lifecycle and a lesson that the RNA genome sequencing must be rigorously validated by cDNA-based phage meeting methods.

RNAcDNA hybrids mediate transposition by way of totally different mechanisms

Retrotransposons can signify half of eukaryotic genomes. Retrotransposon dysregulation destabilizes genomes and has been linked to varied human ailments. Rising regulators of retromobility embrace RNA-DNA hybrid-containing constructions generally known as R-loops. Accumulation of those constructions on the transposons of yeast 1 (Ty1) parts has been proven to extend Ty1 retromobility via an unknown mechanism. Right here, by way of a focused genetic display screen, we recognized the rnh1Δ rad27Δ yeast mutant, which lacked each the Ty1 inhibitor Rad27 and the RNA-DNA hybrid suppressor Rnh1.

The mutant exhibited elevated ranges of Ty1 cDNA-associated RNA-DNA hybrids that promoted Ty1 mobility. Furthermore, on this rnh1Δ rad27Δ mutant, however not within the double RNase H mutant rnh1Δ rnh201Δ, RNA-DNA hybrids preferentially existed as duplex nucleic acid constructions and elevated Ty1 mobility in a Rad52-dependent method.

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

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

Genorise® Canine IL-7 cDNA Clone, 10 ug

GR159002 10 µg
EUR 153

POLR3F siRNA

20-abx929231
  • EUR 661.20
  • EUR 878.40
  • 15 nmol
  • 30 nmol

POLR3F siRNA

20-abx929232
  • EUR 661.20
  • EUR 878.40
  • 15 nmol
  • 30 nmol

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

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

POLR3F Peptide

4727P 0.05 mg
EUR 197.7
Description: (NT) POLR3F peptide

Genorise® Human NT-proBNP cDNA Clone, 10 ug

GR154014 10 µg
EUR 153

POLR3F Antibody

24726-100ul 100ul
EUR 468

POLR3F antibody

70R-2962 50 ug
EUR 560.4
Description: Rabbit polyclonal POLR3F antibody raised against the N terminal of POLR3F

POLR3F antibody

70R-19415 50 ul
EUR 522
Description: Rabbit polyclonal POLR3F antibody

POLR3F antibody

70R-2038 50 ug
EUR 560.4
Description: Rabbit polyclonal POLR3F antibody raised against the middle region of POLR3F

POLR3F antibody

70R-50816 100 ul
EUR 292.8
Description: Purified Polyclonal POLR3F antibody

POLR3F Antibody

4727-002mg 0.02 mg
EUR 206.18
Description: POLR3F Antibody: The human POLR3F is a component of RNA III polymerase. RNA polymerase III transcribes many essential, small, noncoding RNAs, including the 5S rRNAs and tRNAs. While most pol III-transcribed genes are found scattered throughout the linear chromosome maps or in multiple linear clusters, there is increasing evidence that many of these genes prefer to be spatially clustered, often at or near the nucleolus. This association could create an environment that fosters the coregulation of transcription by pol III with transcription of the large ribosomal RNA repeats by RNA polymerase I (pol I) within the nucleolus. Given the high number of pol III-transcribed genes in all eukaryotic genomes, the spatial organization of these genes is likely to affect a large portion of the other genes in a genome. POLR3F has also been recently identified as an HIV dependency factor (HDF), suggesting that POLR3F may be an important drug target in HIV treatment. At least two isoforms of POLR3F are known to exist.

POLR3F Antibody

4727-01mg 0.1 mg
EUR 523.7
Description: POLR3F Antibody: The human POLR3F is a component of RNA III polymerase. RNA polymerase III transcribes many essential, small, noncoding RNAs, including the 5S rRNAs and tRNAs. While most pol III-transcribed genes are found scattered throughout the linear chromosome maps or in multiple linear clusters, there is increasing evidence that many of these genes prefer to be spatially clustered, often at or near the nucleolus. This association could create an environment that fosters the coregulation of transcription by pol III with transcription of the large ribosomal RNA repeats by RNA polymerase I (pol I) within the nucleolus. Given the high number of pol III-transcribed genes in all eukaryotic genomes, the spatial organization of these genes is likely to affect a large portion of the other genes in a genome. POLR3F has also been recently identified as an HIV dependency factor (HDF), suggesting that POLR3F may be an important drug target in HIV treatment. At least two isoforms of POLR3F are known to exist.

POLR3F Antibody

1-CSB-PA884440LA01HU
  • EUR 380.40
  • EUR 402.00
  • 100ug
  • 50ug
Description: A polyclonal antibody against POLR3F. Recognizes POLR3F from Human. This antibody is Unconjugated. Tested in the following application: ELISA, WB; Recommended dilution: WB:1:500-1:2000

POLR3F Antibody

1-CSB-PA018348GA01HU
  • EUR 716.40
  • EUR 399.60
  • 150ul
  • 50ul
Description: A polyclonal antibody against POLR3F. Recognizes POLR3F from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, WB

POLR3F Antibody

E94441 100μg
EUR 255
Description: Available in various conjugation types.

POLR3F Antibody

E38PA2497 100ul
EUR 225
Description: Available in various conjugation types.

POLR3F Antibody

E912224 100ul
EUR 255
Description: Available in various conjugation types.

POLR3F Rabbit pAb

A12224-100ul 100 ul
EUR 369.6

POLR3F Rabbit pAb

A12224-200ul 200 ul
EUR 550.8

POLR3F Rabbit pAb

A12224-20ul 20 ul
EUR 219.6

POLR3F Rabbit pAb

A12224-50ul 50 ul
EUR 267.6

POLR3F Rabbit pAb

E2510867 100ul
EUR 225
Description: Available in various conjugation types.

anti- POLR3F antibody

FNab06632 100µg
EUR 658.5
Description: Antibody raised against POLR3F

POLR3F Blocking Peptide

20-abx063691
  • EUR 326.40
  • EUR 493.20
  • 1 mg
  • 5 mg

POLR3F Blocking Peptide

33R-5238 100 ug
EUR 216
Description: A synthetic peptide for use as a blocking control in assays to test for specificity of POLR3F antibody, catalog no. 70R-2038

POLR3F Blocking Peptide

33R-5647 100 ug
EUR 216
Description: A synthetic peptide for use as a blocking control in assays to test for specificity of POLR3F antibody, catalog no. 70R-2962

POLR3F Polyclonal Antibody

27655-100ul 100ul
EUR 302.4

POLR3F Polyclonal Antibody

27655-50ul 50ul
EUR 224.4

POLR3F Polyclonal Antibody

A72143
  • EUR 302.50
  • EUR 423.50
  • 50 ul
  • 100 ul

POLR3F Polyclonal Antibody

A60346
  • EUR 684.66
  • EUR 117.70
  • EUR 302.50
  • EUR 423.50
  • 100 µg
  • 20 ul
  • 50 ul
  • 100 ul

POLR3F protein (His tag)

30R-2961 100 ug
EUR 386.4
Description: Purified recombinant Human POLR3F protein (His tag)

POLR3F ELISA KIT|Human

EF001942 96 Tests
EUR 826.8

Human POLR3F ELISA KIT

ELI-53168h 96 Tests
EUR 988.8

Mouse Polr3f ELISA KIT

ELI-14354m 96 Tests
EUR 1038

Bovine POLR3F ELISA KIT

ELI-14353b 96 Tests
EUR 1113.6

Human POLR3F shRNA Plasmid

20-abx957233
  • EUR 961.20
  • EUR 1345.20
  • 150 µg
  • 300 µg

Mouse POLR3F shRNA Plasmid

20-abx977203
  • EUR 961.20
  • EUR 1345.20
  • 150 µg
  • 300 µg

POLR3F Antibody, HRP conjugated

1-CSB-PA884440LB01HU
  • EUR 380.40
  • EUR 402.00
  • 100ug
  • 50ug
Description: A polyclonal antibody against POLR3F. Recognizes POLR3F from Human. This antibody is HRP conjugated. Tested in the following application: ELISA

POLR3F Recombinant Protein (Rat)

RP221387 100 ug Ask for price

POLR3F Antibody, FITC conjugated

1-CSB-PA884440LC01HU
  • EUR 380.40
  • EUR 402.00
  • 100ug
  • 50ug
Description: A polyclonal antibody against POLR3F. Recognizes POLR3F from Human. This antibody is FITC conjugated. Tested in the following application: ELISA

POLR3F Recombinant Protein (Mouse)

RP163421 100 ug Ask for price

POLR3F Recombinant Protein (Human)

RP024124 100 ug Ask for price

POLR3F Antibody, Biotin conjugated

1-CSB-PA884440LD01HU
  • EUR 380.40
  • EUR 402.00
  • 100ug
  • 50ug
Description: A polyclonal antibody against POLR3F. Recognizes POLR3F from Human. This antibody is Biotin conjugated. Tested in the following application: ELISA

Polr3f ORF Vector (Rat) (pORF)

ORF073797 1.0 ug DNA
EUR 607.2

POLR3F ORF Vector (Human) (pORF)

ORF008042 1.0 ug DNA
EUR 114

Polr3f ORF Vector (Mouse) (pORF)

ORF054475 1.0 ug DNA
EUR 607.2

POLR3F Polyclonal Conjugated Antibody

C27655 100ul
EUR 476.4

Human POLR3F Protein Lysate 20ug

IHUPOLR3FPLLY20UG each
EUR 213
Description: Human POLR3F Protein Lysate 20ug

POLR3F Polyclonal Antibody, HRP Conjugated

A60349
  • EUR 684.66
  • EUR 302.50
  • EUR 423.50
  • 100 µg
  • 50 ul
  • 100 ul

POLR3F Polyclonal Antibody, FITC Conjugated

A60348
  • EUR 684.66
  • EUR 302.50
  • EUR 423.50
  • 100 µg
  • 50 ul
  • 100 ul

POLR3F Polyclonal Antibody, Biotin Conjugated

A60347
  • EUR 684.66
  • EUR 302.50
  • EUR 423.50
  • 100 µg
  • 50 ul
  • 100 ul

Polr3f sgRNA CRISPR Lentivector set (Rat)

K6496101 3 x 1.0 ug
EUR 406.8

Polr3f 3'UTR GFP Stable Cell Line

TU266543 1.0 ml Ask for price

Polr3f 3'UTR GFP Stable Cell Line

TU166697 1.0 ml Ask for price

POLR3F 3'UTR GFP Stable Cell Line

TU068441 1.0 ml
EUR 1672.8

POLR3F sgRNA CRISPR Lentivector set (Human)

K1685001 3 x 1.0 ug
EUR 406.8

Polr3f sgRNA CRISPR Lentivector set (Mouse)

K4059301 3 x 1.0 ug
EUR 406.8

POLR3F Protein Vector (Rat) (pPM-C-HA)

PV295188 500 ng
EUR 723.6

POLR3F Protein Vector (Rat) (pPB-C-His)

PV295186 500 ng
EUR 723.6

POLR3F Protein Vector (Rat) (pPB-N-His)

PV295187 500 ng
EUR 723.6

POLR3F Protein Vector (Rat) (pPM-C-His)

PV295189 500 ng
EUR 723.6

POLR3F Protein Vector (Human) (pPM-C-HA)

PV032167 500 ng
EUR 394.8

POLR3F Protein Vector (Mouse) (pPM-C-HA)

PV217900 500 ng
EUR 723.6

POLR3F Protein Vector (Human) (pPB-C-His)

PV032165 500 ng
EUR 394.8

POLR3F Protein Vector (Human) (pPB-N-His)

PV032166 500 ng
EUR 394.8

POLR3F Protein Vector (Human) (pPM-C-His)

PV032168 500 ng
EUR 394.8

POLR3F Protein Vector (Mouse) (pPB-C-His)

PV217898 500 ng
EUR 723.6

POLR3F Protein Vector (Mouse) (pPB-N-His)

PV217899 500 ng
EUR 723.6

POLR3F Protein Vector (Mouse) (pPM-C-His)

PV217901 500 ng
EUR 723.6

Polr3f 3'UTR Luciferase Stable Cell Line

TU216543 1.0 ml Ask for price

Polr3f 3'UTR Luciferase Stable Cell Line

TU116697 1.0 ml Ask for price

POLR3F 3'UTR Luciferase Stable Cell Line

TU018441 1.0 ml
EUR 1672.8

RNA Polymerase III Subunit F (POLR3F) Antibody

abx236632-100ug 100 ug
EUR 610.8

RNA Polymerase III Subunit F (POLR3F) Antibody

20-abx114612
  • EUR 878.40
  • EUR 477.60
  • 150 ul
  • 50 ul

RNA Polymerase III Subunit F (POLR3F) Antibody

20-abx126392
  • EUR 493.20
  • EUR 710.40
  • EUR 218.40
  • EUR 376.80
  • 100 ul
  • 200 ul
  • 20 ul
  • 50 ul

RNA Polymerase III Subunit F (POLR3F) Antibody

20-abx008320
  • EUR 360.00
  • EUR 526.80
  • EUR 226.80
  • 100 ul
  • 200 ul
  • 30 ul

RNA Polymerase III Subunit F (POLR3F) Antibody

20-abx302688
  • EUR 493.20
  • EUR 2214.00
  • EUR 718.80
  • EUR 218.40
  • EUR 360.00
  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug

Polr3f sgRNA CRISPR Lentivector (Rat) (Target 1)

K6496102 1.0 ug DNA
EUR 184.8

Polr3f sgRNA CRISPR Lentivector (Rat) (Target 2)

K6496103 1.0 ug DNA
EUR 184.8

Polr3f sgRNA CRISPR Lentivector (Rat) (Target 3)

K6496104 1.0 ug DNA
EUR 184.8

POLR3F sgRNA CRISPR Lentivector (Human) (Target 1)

K1685002 1.0 ug DNA
EUR 184.8

POLR3F sgRNA CRISPR Lentivector (Human) (Target 2)

K1685003 1.0 ug DNA
EUR 184.8

POLR3F sgRNA CRISPR Lentivector (Human) (Target 3)

K1685004 1.0 ug DNA
EUR 184.8

Polr3f sgRNA CRISPR Lentivector (Mouse) (Target 1)

K4059302 1.0 ug DNA
EUR 184.8

Polr3f sgRNA CRISPR Lentivector (Mouse) (Target 2)

K4059303 1.0 ug DNA
EUR 184.8

Polr3f sgRNA CRISPR Lentivector (Mouse) (Target 3)

K4059304 1.0 ug DNA
EUR 184.8

Recombinant Human POLR3F Protein, His, E.coli-1mg

QP13106-1mg 1mg
EUR 3308.4

Recombinant Human POLR3F Protein, His, E.coli-5ug

QP13106-5ug 5ug
EUR 186

Recombinant Human POLR3F Protein, His, E.coli-20ug

QP13106-20ug 20ug
EUR 241.2

RNA Polymerase III Subunit F (POLR3F) Antibody (HRP)

20-abx311789
  • EUR 493.20
  • EUR 2214.00
  • EUR 718.80
  • EUR 218.40
  • EUR 360.00
  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug

RNA Polymerase III Subunit F (POLR3F) Antibody (FITC)

20-abx311790
  • EUR 493.20
  • EUR 2214.00
  • EUR 718.80
  • EUR 218.40
  • EUR 360.00
  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug

RNA Polymerase III Subunit F (POLR3F) Antibody (Biotin)

20-abx311791
  • EUR 493.20
  • EUR 2214.00
  • EUR 718.80
  • EUR 218.40
  • EUR 360.00
  • 100 ug
  • 1 mg
  • 200 ug
  • 20 ug
  • 50 ug

Human RNA Polymerase III Subunit F (POLR3F) ELISA Kit

abx382365-96tests 96 tests
EUR 1093.2

pCDH-CMV-MCS-EF1-Puro cDNA Vector cDNA Clon/Exp Vector [pre-packaged virus]

CD510VB-1 >1 x 10^6 IFUs
EUR 720

POLR3F Polymerase III Polypeptide F Human Recombinant Protein

PROTQ9H1D9 Regular: 20ug
EUR 380.4
Description: POLR3F Human Recombinant produced in E.coli is a single, non-glycosylated polypeptide chain containing 339 amino acids (1-316) and having a molecular mass of 38.1kDa.;POLR3F is fused to a 23 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques.

pCDF1-MCS1 cDNA Cloning and Expression Vector

CD100A-1 10 ug
EUR 483

Polr3f sgRNA CRISPR/Cas9 All-in-One Lentivector set (Rat)

K6496105 3 x 1.0 ug
EUR 451.2

The info point out that in cells missing RNA-DNA hybrid and Ty1 repressors, elevated ranges of RNA-cDNA hybrids, that are related to duplex nucleic acid constructions, increase Ty1 mobility by way of a Rad52-dependent mechanism. In distinction, in cells missing RNA-DNA hybrid repressors alone, elevated ranges of RNA-cDNA hybrids, that are related to triplex nucleic acid constructions, increase Ty1 mobility by way of a Rad52-independent course of. We suggest that duplex and triplex RNA-DNA hybrids promote transposon mobility by way of Rad52-dependent or -independent mechanisms.

Leave a Reply

Your email address will not be published. Required fields are marked *

Related Post