Target-capture full-length double-strand cDNA sequencing for alternative splicing analysis

Target-capture full-length double-strand cDNA sequencing for alternative splicing analysis

Different splicing is a regulated course of by which eukaryotic genes could produce numerous organic merchandise. Defects within the course of sometimes have an effect on mobile operate and may result in illness. Subsequent-generation sequencing (NGS) applied sciences have been developed to detect different splicing occasions; nevertheless, the choice splicing occasions detected by commonplace RNA-Seq could or is probably not derived from full-length RNA. The SMARTer methodology supplies full-length double-strand cDNA synthesis, and the ensuing gene expression patterns correlate strongly with commonplace RNA-Seq.
Nonetheless, it additionally yields non-specific genomic DNA amplification. We improved the SMARTer methodology by using a target-capture full-length double-strand cDNA sequencing methodology. Excessive-fidelity, full-length cDNA is generated by the SMARTer methodology, adopted by target-specific seize with exon probes. The expression sample noticed with this SMARTer Seize methodology was extremely correlated with the outcomes of the unique SMARTer methodology. The quantity and accuracy of the detected splicing occasions had been elevated by eliminating non-specific genomic DNA amplification by the SMARTer Seize.
In comparison with the unique SMARTer methodology, the SMARTer Seize offered 4-fold better detection of different splicing occasions on the identical learn quantity, and it took lower than 1/100 of learn quantity to detect the identical variety of splicing occasions. The % splicing in index (PSI) of the SMARTer Seize is very correlated with the PSI of the SMARTer. These outcomes point out that the SMARTer Seize represents an enchancment of the SMARTer methodology to precisely characterize different splicing repertories in focused genes with out biases.

First report of cDNA clone-launched an infection of maize vegetation with the polerovirus maize yellow mosaic virus (MaYMV)

An East African isolate of the maize-associated polerovirus, maize yellow mosaic virus (MaYMV) was beforehand proven to trigger leaf reddening on singly contaminated maize vegetation (Zea mays). Right here we describe the development of a full-length infectious clone of an East African isolate and, for the primary time, present infectivity of clone-derived transcripts within the main host, maize, via vascular puncture inoculation (VPI), in addition to within the dicot analysis mannequin plant species, Nicotiana benthamiana, via agrobacterium inoculation.
Attribute leaf reddening signs had been noticed in a subset of maize vegetation inoculated with clone-derived transcripts, and an infection was confirmed by RT-PCR and Northern blot analyses. In N. benthamiana vegetation, infections had been completely asymptomatic even at excessive virus titers, as was additionally reported for the cloned Chinese language isolate.
On this examine, nevertheless, we demonstrated that N. benthamiana can function a clone launching platform for maize an infection, as VPI of sap of contaminated N. benthamiana vegetation into maize kernels resulted in an infection and the standard crimson leaf signs. We additional demonstrated that the cloned East African isolate virus was aphid transmissible to maize, with experimental transmission charges as much as 97%, akin to that proven beforehand for the native virus.
Curiously, our knowledge moreover confirmed a definitive correlation of leaf reddening signs with elevated expression of chalcone synthase, thus suggesting upregulation of the flavonoid biosynthesis pathway because the molecular foundation for symptom induction in maize. As the primary report of experimental an infection of maize with transcripts from a cloned polerovirus, this work constitutes a breakthrough for research on molecular maize-polerovirus-aphid interactions.

Building and characterization of an infectious cDNA clone of enterovirus 71: a speedy methodology for rescuing infectious virus primarily based on steady cells expressing T7 polymerase

Enterovirus 71 (EV71) is a causative agent of hand, foot and, mouth illness (HFMD) in younger kids. It’s worthwhile for virologists to develop a quick methodology to rescue infectious virus from a viral cDNA clone. Right here, we report a way for speedy rescue of infectious EV71 by utilizing cells expressing T7 polymerase.
The total-length EV71 genome was amplified in a single step by long-distance PCR with a T7 promoter on the 5′ finish, and the T7 polymerase gene was cloned right into a lentivirus vector for building of a steady cell line expressing T7 polymerase. The infectious virus was quickly and effectively rescued by single transfection of cells with the infectious cDNA clone.
Additional experiments confirmed that the rescued virus had traits much like these of the parental virus. This methodology circumvented the problem in performing in vitro transcription of a protracted linear DNA to acquire high-quality RNA. The development of the viral cDNA clone and the quick rescue of the infectious virus will vastly profit future investigations.
Target-capture full-length double-strand cDNA sequencing for alternative splicing analysis

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. Thus far, no info has been reliably derived relating to 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 website mismatch, and 35,414 unigenes had 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 alerts in L. kaempferi. A lot of DEGs responding to drought stress had 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.

Whole Human Serum antibody

20-S1110G000-V0 10 ml
EUR 133
Description: Goat polyclonal Whole Human Serum antibody

CMV protein (whole cell)

30-AC70 1 ml
EUR 403
Description: Purified native CMV protein (whole cell)

Monkey IgM (whole molecule)

31R-AI093 250 µg
EUR 417
Description: Monkey whole IgM

Goat Anti-human Chorionic Gonadotropin Whole (hCG-Whole) IgG aff pure

HCG14-A 1 mg
EUR 286

cDNA Synthesis SuperMix

20-abx09801420ulSystems
  • EUR 565.00
  • EUR 481.00
  • 100 rxns × 20 ul Systems
  • 50 rxns × 20 ul Systems

Novo? cDNA Kit

M1165-100
EUR 354

Novo? cDNA Kit

M1165-25
EUR 267

Evo? cDNA Supermix

M1168-100
EUR 381

Evo? cDNA Supermix

M1168-25
EUR 267

Novo? cDNA Supermix

M1169-100
EUR 441

Novo? cDNA Supermix

M1169-25
EUR 289

HeLa Cell Lysate (Whole Cell)

LYSATE0001 200ug
EUR 150
Description: This whole cell lysate is prepared from Hela cells using Boster's RIPA Lysis Buffer (AR0105) using a standard whole cell lysate protocol. The concentration was determined using the BCA assay process and then diluted using Dithiothreitol (DTT) and a reducing SDS sample loading buffer, heated for 5 minutes at 100˚C.

Human Hela Whole Cell Lysate

LYSATE0023 200ug
EUR 150
Description: This cell lysate is prepared from human hela using Boster's RIPA Lysis Buffer (AR0105) using a standard whole cell lysate protocol. The concentration was determined using the BCA assay process and then diluted using Dithiothreitol (DTT) and a reducing SDS sample loading buffer, heated for 5 minutes at 100˚C.

Human A549 Whole Cell Lysate

LYSATE0025 200ug
EUR 150
Description: This cell lysate is prepared from human A549 using Boster's RIPA Lysis Buffer (AR0105) using a standard whole cell lysate protocol. The concentration was determined using the BCA assay process and then diluted using Dithiothreitol (DTT) and a reducing SDS sample loading buffer, heated for 5 minutes at 100˚C.

Human Colo320 Whole Cell Lysate

LYSATE0026 200ug
EUR 150
Description: This cell lysate is prepared from human Colo320 using Boster's RIPA Lysis Buffer (AR0105) using a standard whole cell lysate protocol. The concentration was determined using the BCA assay process and then diluted using Dithiothreitol (DTT) and a reducing SDS sample loading buffer, heated for 5 minutes at 100˚C.

Human Jurkat Whole Cell Lysate

LYSATE0027 200ug
EUR 150
Description: This cell lysate is prepared from human Jurkat using Boster's RIPA Lysis Buffer (AR0105) using a standard whole cell lysate protocol. The concentration was determined using the BCA assay process and then diluted using Dithiothreitol (DTT) and a reducing SDS sample loading buffer, heated for 5 minutes at 100˚C.

Human SW620 Whole Cell Lysate

LYSATE0028 200ug
EUR 150
Description: This cell lysate is prepared from human SW620 using Boster's RIPA Lysis Buffer (AR0105) using a standard whole cell lysate protocol. The concentration was determined using the BCA assay process and then diluted using Dithiothreitol (DTT) and a reducing SDS sample loading buffer, heated for 5 minutes at 100˚C.

Human HepG2 Whole Cell Lysate

LYSATE0029 200ug
EUR 150
Description: This cell lysate is prepared from human HepG2 using Boster's RIPA Lysis Buffer (AR0105) using a standard whole cell lysate protocol. The concentration was determined using the BCA assay process and then diluted using Dithiothreitol (DTT) and a reducing SDS sample loading buffer, heated for 5 minutes at 100˚C.

Human K562 Whole Cell Lysate

LYSATE0031 200ug
EUR 150
Description: This cell lysate is prepared from human K562 using Boster's RIPA Lysis Buffer (AR0105) using a standard whole cell lysate protocol. The concentration was determined using the BCA assay process and then diluted using Dithiothreitol (DTT) and a reducing SDS sample loading buffer, heated for 5 minutes at 100˚C.

Human 293T Whole Cell Lysate

LYSATE0032 200ug
EUR 150
Description: This cell lysate is prepared from human 293T using Boster's RIPA Lysis Buffer (AR0105) using a standard whole cell lysate protocol. The concentration was determined using the BCA assay process and then diluted using Dithiothreitol (DTT) and a reducing SDS sample loading buffer, heated for 5 minutes at 100˚C.

Human 22RV1 Whole Cell Lysate

LYSATE0033 200ug
EUR 150
Description: This cell lysate is prepared from human 22RV1 using Boster's RIPA Lysis Buffer (AR0105) using a standard whole cell lysate protocol. The concentration was determined using the BCA assay process and then diluted using Dithiothreitol (DTT) and a reducing SDS sample loading buffer, heated for 5 minutes at 100˚C.

Human U2OS Whole Cell Lysate

LYSATE0034 200ug
EUR 150
Description: This cell lysate is prepared from human U2OS using Boster's RIPA Lysis Buffer (AR0105) using a standard whole cell lysate protocol. The concentration was determined using the BCA assay process and then diluted using Dithiothreitol (DTT) and a reducing SDS sample loading buffer, heated for 5 minutes at 100˚C.

Rat RH35 Whole Cell Lysate

LYSATE0036 200ug
EUR 150
Description: This cell lysate is prepared from rat RH35 using Boster's RIPA Lysis Buffer (AR0105) using a standard whole cell lysate protocol. The concentration was determined using the BCA assay process and then diluted using Dithiothreitol (DTT) and a reducing SDS sample loading buffer, heated for 5 minutes at 100˚C.

Rat PC12 Whole Cell Lysate

LYSATE0037 200ug
EUR 150
Description: This cell lysate is prepared from rat PC12 using Boster's RIPA Lysis Buffer (AR0105) using a standard whole cell lysate protocol. The concentration was determined using the BCA assay process and then diluted using Dithiothreitol (DTT) and a reducing SDS sample loading buffer, heated for 5 minutes at 100˚C.

EpiQuik Whole Cell Extraction Kit

OP-0003 100 Extractions
EUR 270.4
Description: Ask the seller for details

Clearview IM Whole Blood test

CV506815 1 box
EUR 106.98
Description: Please check the datasheet of Clearview IM Whole Blood test before using the test.

Rabbit WHOLE EYE 50 EA

41211-2 Each
EUR 318.39

Rabbit KIDNEY WHOLE 50 EA

41220-2 50 Each
EUR 195.5

Rabbit PITUITARY WHOLE 100 EA*

41233-2 100 EA
EUR 688.14

CK KIDNEY WHOLE 25 EA*

43020-2 25 EA
EUR 240.09

CK WHOLE PITUITARY 25 EA*

43033-2 25 EA
EUR 410.83

Bovine EYE WHOLE 25 EA*

57111-2 25 EA
EUR 295.55

Bovine KIDNEY WHOLE 2 EA*

57120-2 2 EA
EUR 174.84

Bovine PITUITARY WHOLE 25 EA*

57133-2 25 EA
EUR 597.88

Pig WHOLE KIDNEY 10 EA*

59420-2 10 EA
EUR 237.91

Pig PITUITARY WHOLE 25 EA*

59433-2 25 EA
EUR 353.19

Rat Heart Whole tissue lysate

RAL-1461 1 mg
EUR 524

Rat Lung Whole tissue lysate

RAL-1462 1 mg
EUR 524

Rat Brain Whole tissue lysate

RAL-1463 1 mg
EUR 524

Rat Liver Whole tissue lysate

RAL-1464 1 mg
EUR 524

Rat Kidney Whole tissue lysate

RAL-1465 1 mg
EUR 524

Rat Spleen Whole tissue lysate

RAL-1466 1 mg
EUR 524

Rat Pancreas Whole tissue lysate

RAL-1469 1 mg
EUR 524

Rat Adrenal Whole tissue lysate

RAL-1470 100ug
EUR 213

Rat Thymus Whole tissue lysate

RAL-1471 1 mg
EUR 524

Rat Colon Whole tissue lysate

RAL-1472 1 mg
EUR 524

Rat Cerebellum Whole tissue lysate

RAL-1473 100ug
EUR 213

Rat Cerebrum Whole tissue lysate

RAL-1474 100ug
EUR 213

Rat Stomach Whole tissue lysate

RAL-1475 1 mg
EUR 524

Rat Testis Whole tissue lysate

RAL-1476 1 mg
EUR 524

Rat Adipose Whole tissue lysate

RAL-1477 1 mg
EUR 524

Rat Bladder Whole tissue lysate

RAL-1478 1 mg
EUR 524

Rat Eye Whole tissue lysate

RAL-1479 100ug
EUR 213

Rat Skin Whole tissue lysate

RAL-1480 100ug
EUR 213

Arabidopsis Thaliana Whole Tissue Lysate

PABL-1300 50 ug
EUR 164

Tomato Plant Whole Tissue Lysate

PABL-1301 50 ug
EUR 164

Barley Plant Whole Tissue Lysate

PABL-1302 50 ug
EUR 164

Beet Plant Whole Tissue Lysate

PABL-1303 50 ug
EUR 164

Corn Plant Whole Tissue Lysate

PABL-1304 50 ug
EUR 164

Cotton Plant Whole Tissue Lysate

PABL-1305 50 ug
EUR 164

Oat Plant Whole Tissue Lysate

PABL-1307 50 ug
EUR 164

Spinach Plant Whole Tissue Lysate

PABL-1308 50 ug
EUR 164

Sunflower Plant Whole Tissue Lysate

PABL-1309 50 ug
EUR 164

Wheat Plant Whole Tissue Lysate

PABL-1310 50 ug
EUR 164

Mouse Heart Whole tissue lysate

MAL-1401 1 mg
EUR 524

Mouse Lung Whole tissue lysate

MAL-1402 1 mg
EUR 524

Mouse Brain Whole tissue lysate

MAL-1403 1 mg
EUR 524

Mouse Liver Whole tissue lysate

MAL-1404 1 mg
EUR 524

Mouse Kidney Whole tissue lysate

MAL-1405 1 mg
EUR 524

Mouse Spleen Whole tissue lysate

MAL-1406 1 mg
EUR 524

Mouse Thymus Whole tissue lysate

MAL-1409 1 mg
EUR 524

Mouse Bladder Whole tissue lysate

MAL-1410 1 mg
EUR 773

Mouse Colon Whole tissue lysate

MAL-1411 1 mg
EUR 524

Mouse Cerebellum Whole tissue lysate

MAL-1412 1 mg
EUR 524

Mouse Cerebrum Whole tissue lysate

MAL-1413 100ug
EUR 213

Mouse Pancreas Whole tissue lysate

MAL-1414 1 mg
EUR 524

Mouse Stomach Whole tissue lysate

MAL-1415 1 mg
EUR 524

Mouse Testis Whole tissue lysate

MAL-1416 1 mg
EUR 524

Mouse Adrenal Whole tissue lysate

MAL-1417 100ug
EUR 213

Mouse Adipose Whole tissue lysate

MAL-1418 1 mg
EUR 524

Mouse Skin Whole tissue lysate

MAL-1419 100ug
EUR 213

Mouse Eye Whole tissue lysate

MAL-1420 100ug
EUR 213

QuantiChrom Whole Blood Hb Kit

DWHB-250 250
EUR 394
Description: For quantitative determination of hemoglobin and evaluation of drug effects on hemoglobin metabolism. Method: OD570nm. Samples: Whole blood samples. Species: all. Procedure: 5 min. Size: 250 tests. Detection limit: 0.2 g/dL.

Whole Blood DNA Isolation Kit

K528-100
EUR 376

Whole Blood RNA Extraction Kit

K1012050 1 Kit
EUR 333
Description: Can be used for various proteomics studies in both normal and pathological cases. It is an excellent control and suitable for educational purposes. This product is prepared from whole tissue homogenates and has undergone SDS-PAGE quality control analysis. The protein is stored in a buffer with protease inhibitor cocktail fo prevent degradation.

cDNA from Plant Normal Tissue: cDNA from Plant: Arabidopsis

C1634310 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 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 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 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 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 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
  • EUR 620.00
  • EUR 523.00
  • 100 rxns × 20 ul Systems
  • 50 rxns × 20 ul Systems
We detected 73 transcription elements (TFs) below drought stress, together with AP2/ERF, bZIP, TCP, and MYB. This examine supplies fundamental full sequence assets for L. kaempferi analysis and can assist us to higher perceive the capabilities of drought-resistance genes in L. kaempferi.