The resulted cDNA is single-stranded.

cDNA synthesis by Invitrogen SuperScript Double-stranded cDNA Synthesis Kit using a T7-Oligo(dT) promoter primer from Affymetrix, cRNA amplification using ENZO BioArray HighYield RNA Transcript Labeling Kit, Metal-induced hydrolysis by Affymetrix 5 X Fragmentation Buffer, External controls (spikes)- Control Oligo B2 (3 nM), Eukaryotic Hybridization control (Affymetrix) (Small Sample Labeling Protocol vII, Affymetrix).

Double-Stranded cDNA Synthesis Kit - ZAGENO

Incubate at 14°C for 12 hours and recover double-stranded cDNA by GENECLEAN II.

Invitrogen SuperScript Double-Stranded cDNA Synthesis Kit

In some experiments such as microarray, double-stranded cDNA is required and can be produced by another round of DNA synthesis following the first strand synthesis.

Search Results for double stranded cdna on ..

Heterologous expression of cDNA library in Arabidopsis and other plants has been used for gene identifications. To identify functions of tomato genes, we expressed a tomato full-length cDNA library in Arabidopsis thaliana and generated over 7,000 mutants. We constructed a tomato cDNA library with a plant transformation-ready binary vector that contained a higher percentage of full-length cDNAs since synthesized double-stranded cDNA was size-selected using gel electrophoresis, with cDNA sizes of 2–5 kb being gel-purified for ligation onto the binary vector. Sequencing of 81 cDNA clones indicates that 75% (61) are full-length genes, which is similar to sequencing of inserted cDNA in Arabidopsis. The library was used to transform Arabidopsis plants. Among the 7,000 mutants, one was found to be a dwarf due to the expression of an ATP synthase, and another vegetative mutant did not produce flowers even after 7 months. The technique was validated by reintroducing the tomato ribosomal protein L9 gene and can be used in any other plant species as a gene discovery tool.

Meeting room discussion covers pain points, comparison.Comparing cDNA Synthesis Kits By Ryan Matson.
Use Gene Synthesis to synthesise codon-optimised.RNA Isolation from Embryonic Zebrafish and cDNA Synthesis for Gene Expression Analysis.

Invitrogen™ NCode™ miRNA First-Strand cDNA Synthesis Kit

During first-strand SMARTer cDNA synthesis known universal primer sequences are incorporated at both ends of the cDNA. Following first-strand synthesis, SMARTer cDNA is immediately available for PCR amplification.

Once mRNA is converted to cDNA, the cDNA can be used for RT-PCR, as probe for expression analysis and for cloning of the mRNA sequence.

A strategy for rapid cDNA cloning from double-stranded …

When RNA transcripts synthesized from the full-length CHV1-EP721 cDNA template were used to transfect the virus-free fungal strain EP721(-v), colonies with traits indistinguishable from those of CHV1-EP721-containing strain EP721, including a white phenotype, suppressed sporulation, and the presence of cytoplasmically replicative viral dsRNA, were obtained, demonstrating a successful infection. To further examine whether different fungal hosts would have an impact on the accumulation of viral dsRNA, hypoviruses CHV1-EP713, CHV1-Euro7, and CHV1-EP721 were introduced into virus-free and genetically different fungal strains EP155, Euro7(-v), and EP721(-v), respectively, via transfection. A change of colony pigmentation from orange to white indicated that a productive infection had been established for all these viruses. All three fungal hosts exhibited similar symptoms (white pigmentation and suppression of conidial spore production) upon infection by the same hypovirus, and different virus strains incited subtle yet distinguishable symptoms in these hosts (Fig. ). These results demonstrate that the phenotype of a transfectant is determined predominantly by the virus and not by the host. It is evident from Fig. that morphologies of transfectants infected by CHV1-EP721 are more similar to those of transfectants infected by CHV1-Euro7 than by CHV1-EP713. It was also observed that at a later stage of infection (7 to 10 days), the colony morphology and pigmentation exhibited some differences for the three fungal hosts infected with CHV1-EP721, with hosts of Euro7(-v) and EP721(-v) being whiter and making some stroma-like structures, two characteristics found in native EP721. This could be regarded as the contribution of the host genome to symptom expression (). An examination of viral dsRNA in the infected cells revealed that the lowest dsRNA accumulation was found in the CHV1-EP721-infected strains, and the highest level was found in the CHV1-EP713-infected strains, with CHV1-Euro7 RNA accumulating to a level about half of that of CHV1-EP713 (Fig. ). These results clearly demonstrated that the viral dsRNA accumulation level was determined solely by the input virus.

using 5 µl of the double stranded cDNA, standard Taq polymerase (Invitrogen) ..

second strand for double stranded C-DNA …

Hypoviruses do not encode a capsid protein, and hypovirus RNA replication occurs in association with host membrane vesicles that accumulate viral double-stranded RNA (dsRNA). Like other mycoviruses, hypoviruses lack an extracellular route of infection. Natural virus transmission is restricted to cytoplasmic mixing as a result of fusion (anastomosis) between vegetatively compatible strains or to various degrees through asexual spores (conidia). The genome of the prototypic hypovirus CHV1-EP713 consists of a 12.7-kb coding strand that contains two open reading frames (ORFs), ORF A and ORF B, separated by a UAAUG pentanucleotide (). ORF A encodes a polypeptide, p69, which is autocatalytically processed into two products, p29 and p40, by the action of a papain-like protease domain located in the p29 coding region. Cleavage occurs between p69 Gly248 and Gly249, and the residues Cys162 and His215 are essential for autolytic cleavage (). Expression of ORF A in C. parasitica tranformants in the absence of virus infection resulted in a hypovirus-associated loss of orange pigment production (white phenotype) and a significant reduction in asexual sporulation but not in virulence attenuation (). Craven et al. subsequently mapped the ORF A suppressive function to p29 at the N-terminal portion of p69 ().