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TaKaRa cdna amplification
Features of the <t>RACE</t> products. (a) Connectivity of detected transcripts to known exons/novel transcriptionally active regions (TARs). (b) Frequency of splice and unspliced rapid amplification of <t>cDNA</t> ends (RACE) products derived from known exons, novel TARs, and untranscribed regions. (c) Average microarray intensities of regions encoding spliced and unspliced RACE products. nontx, region not previously shown to be transcribed.
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1) Product Images from "Systematic analysis of transcribed loci in ENCODE regions using RACE sequencing reveals extensive transcription in the human genome"

Article Title: Systematic analysis of transcribed loci in ENCODE regions using RACE sequencing reveals extensive transcription in the human genome

Journal: Genome Biology

doi: 10.1186/gb-2008-9-1-r3

Features of the RACE products. (a) Connectivity of detected transcripts to known exons/novel transcriptionally active regions (TARs). (b) Frequency of splice and unspliced rapid amplification of cDNA ends (RACE) products derived from known exons, novel TARs, and untranscribed regions. (c) Average microarray intensities of regions encoding spliced and unspliced RACE products. nontx, region not previously shown to be transcribed.
Figure Legend Snippet: Features of the RACE products. (a) Connectivity of detected transcripts to known exons/novel transcriptionally active regions (TARs). (b) Frequency of splice and unspliced rapid amplification of cDNA ends (RACE) products derived from known exons, novel TARs, and untranscribed regions. (c) Average microarray intensities of regions encoding spliced and unspliced RACE products. nontx, region not previously shown to be transcribed.

Techniques Used: Rapid Amplification of cDNA Ends, Derivative Assay, Microarray

Distribution of RACE product sequences in the DRG1 and FBX07 regions. (a) DRG1 Region and (b) FBX07 region. Products from the sense strand (+) are shown in the top half of the panel. Products from the antisense strand are in the bottom half of the panel. Blue products are detected sequences from 5'-rapid amplification of cDNA ends (RACE); red products are detected sequences from 3'-RACE; black indicates refSeq; black asterisks indicate consensus splice sites (GT-AG, GC-AG, or AT-AC); and green asterisks indicate novel isoforms with more than 50% consensus splice sites. Note that the antisense products that lack consensus splice sites are indicated in lighter colors. (c) cDNA and RNA hybridization signals in DRG1 region. The blue tracks indicate the signals that were generated from hybridization of cDNA prepared from NB4 cells using reverse transcriptase to the strand-specific microarray. The red tracks indicate hybridization of RNA that has been labeled directly by chemical means, thus omitting the use of reverse transcriptase, to the strand-specific microarray. Products from the sense strand (+) are shown in the top half of the panel. Products from the antisense strand are in the bottom half of the panel.
Figure Legend Snippet: Distribution of RACE product sequences in the DRG1 and FBX07 regions. (a) DRG1 Region and (b) FBX07 region. Products from the sense strand (+) are shown in the top half of the panel. Products from the antisense strand are in the bottom half of the panel. Blue products are detected sequences from 5'-rapid amplification of cDNA ends (RACE); red products are detected sequences from 3'-RACE; black indicates refSeq; black asterisks indicate consensus splice sites (GT-AG, GC-AG, or AT-AC); and green asterisks indicate novel isoforms with more than 50% consensus splice sites. Note that the antisense products that lack consensus splice sites are indicated in lighter colors. (c) cDNA and RNA hybridization signals in DRG1 region. The blue tracks indicate the signals that were generated from hybridization of cDNA prepared from NB4 cells using reverse transcriptase to the strand-specific microarray. The red tracks indicate hybridization of RNA that has been labeled directly by chemical means, thus omitting the use of reverse transcriptase, to the strand-specific microarray. Products from the sense strand (+) are shown in the top half of the panel. Products from the antisense strand are in the bottom half of the panel.

Techniques Used: Rapid Amplification of cDNA Ends, Hybridization, Generated, Microarray, Labeling

RACE sequencing can detect transcripts not previously detected by microarray analysis in NB4 cells. (a) Integrated Genome Browser (IGB) view SYN3 and TIMP3 rapid amplification of cDNA ends (RACE) products in NB4 RNA. (b) Real-time PCR quantification of SYN3 and TIMP3 transcripts relative to HPRT1 in NB4 cells.
Figure Legend Snippet: RACE sequencing can detect transcripts not previously detected by microarray analysis in NB4 cells. (a) Integrated Genome Browser (IGB) view SYN3 and TIMP3 rapid amplification of cDNA ends (RACE) products in NB4 RNA. (b) Real-time PCR quantification of SYN3 and TIMP3 transcripts relative to HPRT1 in NB4 cells.

Techniques Used: Sequencing, Microarray, Rapid Amplification of cDNA Ends, Real-time Polymerase Chain Reaction

Example of a novel transcript detected by RACE sequencing. (a) Novel transcript 5NGSP2F8 (with consensus splice site) has a potential open reading frame of 142 amino acids; also, there is spliced expressed sequence tag (EST) evidence for it. (b) Real-time PCR relative quantification of the novel transcript to HPRT1 in placenta polyA+ RNA. RACE, rapid amplification of cDNA ends.
Figure Legend Snippet: Example of a novel transcript detected by RACE sequencing. (a) Novel transcript 5NGSP2F8 (with consensus splice site) has a potential open reading frame of 142 amino acids; also, there is spliced expressed sequence tag (EST) evidence for it. (b) Real-time PCR relative quantification of the novel transcript to HPRT1 in placenta polyA+ RNA. RACE, rapid amplification of cDNA ends.

Techniques Used: Sequencing, Real-time Polymerase Chain Reaction, Rapid Amplification of cDNA Ends

Frequency of PCR products obtained from different genomic regions. Primers designed to the sense and antisense strands of exons, novel transcriptionally active regions (TARs) and nontranscribed regions were used to generate rapid amplification of cDNA ends (RACE) products. The frequency of PCR products obtained is indicated. nontx, region not previously shown to be transcribed.
Figure Legend Snippet: Frequency of PCR products obtained from different genomic regions. Primers designed to the sense and antisense strands of exons, novel transcriptionally active regions (TARs) and nontranscribed regions were used to generate rapid amplification of cDNA ends (RACE) products. The frequency of PCR products obtained is indicated. nontx, region not previously shown to be transcribed.

Techniques Used: Polymerase Chain Reaction, Rapid Amplification of cDNA Ends

2) Product Images from "Array Analysis of Viral Gene Transcription during Lytic Infection of Cells in Tissue Culture with Varicella-Zoster Virus"

Article Title: Array Analysis of Viral Gene Transcription during Lytic Infection of Cells in Tissue Culture with Varicella-Zoster Virus

Journal: Journal of Virology

doi: 10.1128/JVI.77.21.11718-11732.2003

RT-PCR analysis of VZV intergenic DNA. Control (C) and VZV-infected (V) cell RNA was incubated with (+) or without (−) reverse transcriptase and PCR amplified with selected intergenic (Int)-specific primers. Amplified products were compared with those after PCR amplification of VZV DNA. The predicted size of the amplification product (amplicon size) for each Int region primer set is listed. Int regions 6 to 8 map between ORFs 60 and 61; Int regions 10 to 12 map between ORFs 61 and 62 and contain the IR L /IR S junction; and Int regions 1 to 5 map between ORFs 62 and 63 and contain the VZV DNA origin replication (ori).
Figure Legend Snippet: RT-PCR analysis of VZV intergenic DNA. Control (C) and VZV-infected (V) cell RNA was incubated with (+) or without (−) reverse transcriptase and PCR amplified with selected intergenic (Int)-specific primers. Amplified products were compared with those after PCR amplification of VZV DNA. The predicted size of the amplification product (amplicon size) for each Int region primer set is listed. Int regions 6 to 8 map between ORFs 60 and 61; Int regions 10 to 12 map between ORFs 61 and 62 and contain the IR L /IR S junction; and Int regions 1 to 5 map between ORFs 62 and 63 and contain the VZV DNA origin replication (ori).

Techniques Used: Reverse Transcription Polymerase Chain Reaction, Infection, Incubation, Polymerase Chain Reaction, Amplification

3) Product Images from "Systematic analysis of transcribed loci in ENCODE regions using RACE sequencing reveals extensive transcription in the human genome"

Article Title: Systematic analysis of transcribed loci in ENCODE regions using RACE sequencing reveals extensive transcription in the human genome

Journal: Genome Biology

doi: 10.1186/gb-2008-9-1-r3

Features of the RACE products. (a) Connectivity of detected transcripts to known exons/novel transcriptionally active regions (TARs). (b) Frequency of splice and unspliced rapid amplification of cDNA ends (RACE) products derived from known exons, novel TARs, and untranscribed regions. (c) Average microarray intensities of regions encoding spliced and unspliced RACE products. nontx, region not previously shown to be transcribed.
Figure Legend Snippet: Features of the RACE products. (a) Connectivity of detected transcripts to known exons/novel transcriptionally active regions (TARs). (b) Frequency of splice and unspliced rapid amplification of cDNA ends (RACE) products derived from known exons, novel TARs, and untranscribed regions. (c) Average microarray intensities of regions encoding spliced and unspliced RACE products. nontx, region not previously shown to be transcribed.

Techniques Used: Rapid Amplification of cDNA Ends, Derivative Assay, Microarray

Distribution of RACE product sequences in the DRG1 and FBX07 regions. (a) DRG1 Region and (b) FBX07 region. Products from the sense strand (+) are shown in the top half of the panel. Products from the antisense strand are in the bottom half of the panel. Blue products are detected sequences from 5'-rapid amplification of cDNA ends (RACE); red products are detected sequences from 3'-RACE; black indicates refSeq; black asterisks indicate consensus splice sites (GT-AG, GC-AG, or AT-AC); and green asterisks indicate novel isoforms with more than 50% consensus splice sites. Note that the antisense products that lack consensus splice sites are indicated in lighter colors. (c) cDNA and RNA hybridization signals in DRG1 region. The blue tracks indicate the signals that were generated from hybridization of cDNA prepared from NB4 cells using reverse transcriptase to the strand-specific microarray. The red tracks indicate hybridization of RNA that has been labeled directly by chemical means, thus omitting the use of reverse transcriptase, to the strand-specific microarray. Products from the sense strand (+) are shown in the top half of the panel. Products from the antisense strand are in the bottom half of the panel.
Figure Legend Snippet: Distribution of RACE product sequences in the DRG1 and FBX07 regions. (a) DRG1 Region and (b) FBX07 region. Products from the sense strand (+) are shown in the top half of the panel. Products from the antisense strand are in the bottom half of the panel. Blue products are detected sequences from 5'-rapid amplification of cDNA ends (RACE); red products are detected sequences from 3'-RACE; black indicates refSeq; black asterisks indicate consensus splice sites (GT-AG, GC-AG, or AT-AC); and green asterisks indicate novel isoforms with more than 50% consensus splice sites. Note that the antisense products that lack consensus splice sites are indicated in lighter colors. (c) cDNA and RNA hybridization signals in DRG1 region. The blue tracks indicate the signals that were generated from hybridization of cDNA prepared from NB4 cells using reverse transcriptase to the strand-specific microarray. The red tracks indicate hybridization of RNA that has been labeled directly by chemical means, thus omitting the use of reverse transcriptase, to the strand-specific microarray. Products from the sense strand (+) are shown in the top half of the panel. Products from the antisense strand are in the bottom half of the panel.

Techniques Used: Rapid Amplification of cDNA Ends, Hybridization, Generated, Microarray, Labeling

RACE sequencing can detect transcripts not previously detected by microarray analysis in NB4 cells. (a) Integrated Genome Browser (IGB) view SYN3 and TIMP3 rapid amplification of cDNA ends (RACE) products in NB4 RNA. (b) Real-time PCR quantification of SYN3 and TIMP3 transcripts relative to HPRT1 in NB4 cells.
Figure Legend Snippet: RACE sequencing can detect transcripts not previously detected by microarray analysis in NB4 cells. (a) Integrated Genome Browser (IGB) view SYN3 and TIMP3 rapid amplification of cDNA ends (RACE) products in NB4 RNA. (b) Real-time PCR quantification of SYN3 and TIMP3 transcripts relative to HPRT1 in NB4 cells.

Techniques Used: Sequencing, Microarray, Rapid Amplification of cDNA Ends, Real-time Polymerase Chain Reaction

Example of a novel transcript detected by RACE sequencing. (a) Novel transcript 5NGSP2F8 (with consensus splice site) has a potential open reading frame of 142 amino acids; also, there is spliced expressed sequence tag (EST) evidence for it. (b) Real-time PCR relative quantification of the novel transcript to HPRT1 in placenta polyA+ RNA. RACE, rapid amplification of cDNA ends.
Figure Legend Snippet: Example of a novel transcript detected by RACE sequencing. (a) Novel transcript 5NGSP2F8 (with consensus splice site) has a potential open reading frame of 142 amino acids; also, there is spliced expressed sequence tag (EST) evidence for it. (b) Real-time PCR relative quantification of the novel transcript to HPRT1 in placenta polyA+ RNA. RACE, rapid amplification of cDNA ends.

Techniques Used: Sequencing, Real-time Polymerase Chain Reaction, Rapid Amplification of cDNA Ends

Frequency of PCR products obtained from different genomic regions. Primers designed to the sense and antisense strands of exons, novel transcriptionally active regions (TARs) and nontranscribed regions were used to generate rapid amplification of cDNA ends (RACE) products. The frequency of PCR products obtained is indicated. nontx, region not previously shown to be transcribed.
Figure Legend Snippet: Frequency of PCR products obtained from different genomic regions. Primers designed to the sense and antisense strands of exons, novel transcriptionally active regions (TARs) and nontranscribed regions were used to generate rapid amplification of cDNA ends (RACE) products. The frequency of PCR products obtained is indicated. nontx, region not previously shown to be transcribed.

Techniques Used: Polymerase Chain Reaction, Rapid Amplification of cDNA Ends

4) Product Images from "Systematic analysis of transcribed loci in ENCODE regions using RACE sequencing reveals extensive transcription in the human genome"

Article Title: Systematic analysis of transcribed loci in ENCODE regions using RACE sequencing reveals extensive transcription in the human genome

Journal: Genome Biology

doi: 10.1186/gb-2008-9-1-r3

Features of the RACE products. (a) Connectivity of detected transcripts to known exons/novel transcriptionally active regions (TARs). (b) Frequency of splice and unspliced rapid amplification of cDNA ends (RACE) products derived from known exons, novel TARs, and untranscribed regions. (c) Average microarray intensities of regions encoding spliced and unspliced RACE products. nontx, region not previously shown to be transcribed.
Figure Legend Snippet: Features of the RACE products. (a) Connectivity of detected transcripts to known exons/novel transcriptionally active regions (TARs). (b) Frequency of splice and unspliced rapid amplification of cDNA ends (RACE) products derived from known exons, novel TARs, and untranscribed regions. (c) Average microarray intensities of regions encoding spliced and unspliced RACE products. nontx, region not previously shown to be transcribed.

Techniques Used: Rapid Amplification of cDNA Ends, Derivative Assay, Microarray

Distribution of RACE product sequences in the DRG1 and FBX07 regions. (a) DRG1 Region and (b) FBX07 region. Products from the sense strand (+) are shown in the top half of the panel. Products from the antisense strand are in the bottom half of the panel. Blue products are detected sequences from 5'-rapid amplification of cDNA ends (RACE); red products are detected sequences from 3'-RACE; black indicates refSeq; black asterisks indicate consensus splice sites (GT-AG, GC-AG, or AT-AC); and green asterisks indicate novel isoforms with more than 50% consensus splice sites. Note that the antisense products that lack consensus splice sites are indicated in lighter colors. (c) cDNA and RNA hybridization signals in DRG1 region. The blue tracks indicate the signals that were generated from hybridization of cDNA prepared from NB4 cells using reverse transcriptase to the strand-specific microarray. The red tracks indicate hybridization of RNA that has been labeled directly by chemical means, thus omitting the use of reverse transcriptase, to the strand-specific microarray. Products from the sense strand (+) are shown in the top half of the panel. Products from the antisense strand are in the bottom half of the panel.
Figure Legend Snippet: Distribution of RACE product sequences in the DRG1 and FBX07 regions. (a) DRG1 Region and (b) FBX07 region. Products from the sense strand (+) are shown in the top half of the panel. Products from the antisense strand are in the bottom half of the panel. Blue products are detected sequences from 5'-rapid amplification of cDNA ends (RACE); red products are detected sequences from 3'-RACE; black indicates refSeq; black asterisks indicate consensus splice sites (GT-AG, GC-AG, or AT-AC); and green asterisks indicate novel isoforms with more than 50% consensus splice sites. Note that the antisense products that lack consensus splice sites are indicated in lighter colors. (c) cDNA and RNA hybridization signals in DRG1 region. The blue tracks indicate the signals that were generated from hybridization of cDNA prepared from NB4 cells using reverse transcriptase to the strand-specific microarray. The red tracks indicate hybridization of RNA that has been labeled directly by chemical means, thus omitting the use of reverse transcriptase, to the strand-specific microarray. Products from the sense strand (+) are shown in the top half of the panel. Products from the antisense strand are in the bottom half of the panel.

Techniques Used: Rapid Amplification of cDNA Ends, Hybridization, Generated, Microarray, Labeling

RACE sequencing can detect transcripts not previously detected by microarray analysis in NB4 cells. (a) Integrated Genome Browser (IGB) view SYN3 and TIMP3 rapid amplification of cDNA ends (RACE) products in NB4 RNA. (b) Real-time PCR quantification of SYN3 and TIMP3 transcripts relative to HPRT1 in NB4 cells.
Figure Legend Snippet: RACE sequencing can detect transcripts not previously detected by microarray analysis in NB4 cells. (a) Integrated Genome Browser (IGB) view SYN3 and TIMP3 rapid amplification of cDNA ends (RACE) products in NB4 RNA. (b) Real-time PCR quantification of SYN3 and TIMP3 transcripts relative to HPRT1 in NB4 cells.

Techniques Used: Sequencing, Microarray, Rapid Amplification of cDNA Ends, Real-time Polymerase Chain Reaction

Example of a novel transcript detected by RACE sequencing. (a) Novel transcript 5NGSP2F8 (with consensus splice site) has a potential open reading frame of 142 amino acids; also, there is spliced expressed sequence tag (EST) evidence for it. (b) Real-time PCR relative quantification of the novel transcript to HPRT1 in placenta polyA+ RNA. RACE, rapid amplification of cDNA ends.
Figure Legend Snippet: Example of a novel transcript detected by RACE sequencing. (a) Novel transcript 5NGSP2F8 (with consensus splice site) has a potential open reading frame of 142 amino acids; also, there is spliced expressed sequence tag (EST) evidence for it. (b) Real-time PCR relative quantification of the novel transcript to HPRT1 in placenta polyA+ RNA. RACE, rapid amplification of cDNA ends.

Techniques Used: Sequencing, Real-time Polymerase Chain Reaction, Rapid Amplification of cDNA Ends

Frequency of PCR products obtained from different genomic regions. Primers designed to the sense and antisense strands of exons, novel transcriptionally active regions (TARs) and nontranscribed regions were used to generate rapid amplification of cDNA ends (RACE) products. The frequency of PCR products obtained is indicated. nontx, region not previously shown to be transcribed.
Figure Legend Snippet: Frequency of PCR products obtained from different genomic regions. Primers designed to the sense and antisense strands of exons, novel transcriptionally active regions (TARs) and nontranscribed regions were used to generate rapid amplification of cDNA ends (RACE) products. The frequency of PCR products obtained is indicated. nontx, region not previously shown to be transcribed.

Techniques Used: Polymerase Chain Reaction, Rapid Amplification of cDNA Ends

5) Product Images from "Autoinhibitory Regulation of p73 by ?Np73 To Modulate Cell Survival and Death through a p73-Specific Target Element within the ?Np73 Promoter"

Article Title: Autoinhibitory Regulation of p73 by ?Np73 To Modulate Cell Survival and Death through a p73-Specific Target Element within the ?Np73 Promoter

Journal: Molecular and Cellular Biology

doi: 10.1128/MCB.22.8.2575-2585.2002

Induction of ΔNp73 by p73α. (A) Western blot analysis. SH-SY5Y cells were infected with the recombinant adenovirus for LacZ (Ad-LacZ), p53 (Ad-p53), or HA-p73α (Ad-p73α) at an MOI of 10. At 36 h after the infection, whole-cell lysates (40 μg) were prepared and then subjected to Western blotting with anti-p73α (top) or anti-p53 (middle) antibody. The p73α blot was reprobed for actin to ensure equal protein loading (bottom). (B) Northern blot analysis of ΔNp73 expression. SH-SY5Y cells were infected with an MOI of 10 of adenovirus expressing LacZ, p53, or HA-p73α. Total RNA (20 μg) was prepared 24 h postinfection and subjected to Northern blotting with the ΔNp73 -specific cDNA probe. Ethidium bromide staining of 28S and 18S rRNAs is shown to allow comparison of RNA loaded. (C) RT-PCR analysis of HA - p73 or ΔNp73 expression in adenovirus-infected SH-SY5Y, SK-N-AS, or SK-N-BE cells (MOI = 10). GAPDH expression is shown as a control.
Figure Legend Snippet: Induction of ΔNp73 by p73α. (A) Western blot analysis. SH-SY5Y cells were infected with the recombinant adenovirus for LacZ (Ad-LacZ), p53 (Ad-p53), or HA-p73α (Ad-p73α) at an MOI of 10. At 36 h after the infection, whole-cell lysates (40 μg) were prepared and then subjected to Western blotting with anti-p73α (top) or anti-p53 (middle) antibody. The p73α blot was reprobed for actin to ensure equal protein loading (bottom). (B) Northern blot analysis of ΔNp73 expression. SH-SY5Y cells were infected with an MOI of 10 of adenovirus expressing LacZ, p53, or HA-p73α. Total RNA (20 μg) was prepared 24 h postinfection and subjected to Northern blotting with the ΔNp73 -specific cDNA probe. Ethidium bromide staining of 28S and 18S rRNAs is shown to allow comparison of RNA loaded. (C) RT-PCR analysis of HA - p73 or ΔNp73 expression in adenovirus-infected SH-SY5Y, SK-N-AS, or SK-N-BE cells (MOI = 10). GAPDH expression is shown as a control.

Techniques Used: Western Blot, Infection, Recombinant, Northern Blot, Expressing, Staining, Reverse Transcription Polymerase Chain Reaction

Related Articles

Amplification:

Article Title: The pre- vertebrate origins of neurogenic placodes
Article Snippet: .. For the calcium imaging experiments, cDNA fragments containing the full-length coding regions of C. intestinalis CNGA, CNGB and CNGC (Ghost Gene IDs: KH.C2.249,KH.L42.6, and KH.C7.605, respectively) were amplified from a cDNA pool of mid tailbud embryos (CNGA) or larvae (CNGB and CNGC) by PCR using a thermostable DNA polymerase exhibiting proofreading activity (Takara LATaq; Takara Bio) with gene-specific primers (59-AGCTAACACAGATTTTAGTGTAATG-39 and 59-GTTGCGGGTAAATTACATGTC-39 for CNGA; 59-ATGGCATTGCACATAAATGCAA-39 and 59-AGCAAAGACTTTGGTAAACATCAG-39 for CNGB; 59-TAAGGTTGATACAGGTTTCATTGG-39 and 59-GACGTAATCATGACGAAACTGTG-39 for CNGC). .. The PCR products were subcloned into pcDNA6 plasmid vectors and sequenced on both strands using the cycle sequencing method with an Applied Biosystems 3100 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA).

Article Title: Comparative Analysis of Gtf Isozyme Production and Diversity in Isolates of Streptococcus mutans with Different Biofilm Growth Phenotypes
Article Snippet: .. Amplification reactions were performed with a polymerase enzyme system specific for the generation of long PCR products (Advantage2; Clontech Labs, Inc.). ..

Synthesized:

Article Title: MiR-20a Promotes Cervical Cancer Proliferation and Metastasis In Vitro and In Vivo
Article Snippet: .. The reverse-transcription reactions were carried out using an MiraMasTM Kit (Bioo scientific, USA), which contains poly (A) polymerase used for polyadenylation of miRNA. qRT-PCR was performed using a standard SYBR Green PCR kit (Takara, Japan).The primers were synthesized (Shanghai GenePharma, China) as follows: miR-20a forwards primer: TAC GAT AAA GTG CTT ATA GTG CAG GTA G. U6 forwards primer: ATT GGA ACG ATA CAG AGA AGA TT. ..

Quantitative RT-PCR:

Article Title: MiR-20a Promotes Cervical Cancer Proliferation and Metastasis In Vitro and In Vivo
Article Snippet: .. The reverse-transcription reactions were carried out using an MiraMasTM Kit (Bioo scientific, USA), which contains poly (A) polymerase used for polyadenylation of miRNA. qRT-PCR was performed using a standard SYBR Green PCR kit (Takara, Japan).The primers were synthesized (Shanghai GenePharma, China) as follows: miR-20a forwards primer: TAC GAT AAA GTG CTT ATA GTG CAG GTA G. U6 forwards primer: ATT GGA ACG ATA CAG AGA AGA TT. ..

SYBR Green Assay:

Article Title: MiR-20a Promotes Cervical Cancer Proliferation and Metastasis In Vitro and In Vivo
Article Snippet: .. The reverse-transcription reactions were carried out using an MiraMasTM Kit (Bioo scientific, USA), which contains poly (A) polymerase used for polyadenylation of miRNA. qRT-PCR was performed using a standard SYBR Green PCR kit (Takara, Japan).The primers were synthesized (Shanghai GenePharma, China) as follows: miR-20a forwards primer: TAC GAT AAA GTG CTT ATA GTG CAG GTA G. U6 forwards primer: ATT GGA ACG ATA CAG AGA AGA TT. ..

Reverse Transcription Polymerase Chain Reaction:

Article Title: Complete Genome Sequence Analysis of Two Divergent Groups of Sweet potato chlorotic fleck virus Isolates Collected from Korea
Article Snippet: .. Reverse transcription polymerase chain reaction (RT-PCR) was performed as a two-step procedure; RT was conducted using Avian myeloblastosis virus (AMV) reverse transcriptase (Promega, USA), and PCR was carried out using high-fidelity LA taq polymerase (Takara, Japan). .. Specific PCR primer pairs were designed for primer walking and subsequent sequencing to obtain the complete genome sequences based on previously reported SPCFV nucleotide sequences ( , ).

Activity Assay:

Article Title: The pre- vertebrate origins of neurogenic placodes
Article Snippet: .. For the calcium imaging experiments, cDNA fragments containing the full-length coding regions of C. intestinalis CNGA, CNGB and CNGC (Ghost Gene IDs: KH.C2.249,KH.L42.6, and KH.C7.605, respectively) were amplified from a cDNA pool of mid tailbud embryos (CNGA) or larvae (CNGB and CNGC) by PCR using a thermostable DNA polymerase exhibiting proofreading activity (Takara LATaq; Takara Bio) with gene-specific primers (59-AGCTAACACAGATTTTAGTGTAATG-39 and 59-GTTGCGGGTAAATTACATGTC-39 for CNGA; 59-ATGGCATTGCACATAAATGCAA-39 and 59-AGCAAAGACTTTGGTAAACATCAG-39 for CNGB; 59-TAAGGTTGATACAGGTTTCATTGG-39 and 59-GACGTAATCATGACGAAACTGTG-39 for CNGC). .. The PCR products were subcloned into pcDNA6 plasmid vectors and sequenced on both strands using the cycle sequencing method with an Applied Biosystems 3100 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA).

Imaging:

Article Title: The pre- vertebrate origins of neurogenic placodes
Article Snippet: .. For the calcium imaging experiments, cDNA fragments containing the full-length coding regions of C. intestinalis CNGA, CNGB and CNGC (Ghost Gene IDs: KH.C2.249,KH.L42.6, and KH.C7.605, respectively) were amplified from a cDNA pool of mid tailbud embryos (CNGA) or larvae (CNGB and CNGC) by PCR using a thermostable DNA polymerase exhibiting proofreading activity (Takara LATaq; Takara Bio) with gene-specific primers (59-AGCTAACACAGATTTTAGTGTAATG-39 and 59-GTTGCGGGTAAATTACATGTC-39 for CNGA; 59-ATGGCATTGCACATAAATGCAA-39 and 59-AGCAAAGACTTTGGTAAACATCAG-39 for CNGB; 59-TAAGGTTGATACAGGTTTCATTGG-39 and 59-GACGTAATCATGACGAAACTGTG-39 for CNGC). .. The PCR products were subcloned into pcDNA6 plasmid vectors and sequenced on both strands using the cycle sequencing method with an Applied Biosystems 3100 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA).

Expressing:

Article Title: MiR-148b suppresses cell proliferation and invasion in hepatocellular carcinoma by targeting WNT1/β-catenin pathway
Article Snippet: .. To quantify mature miR-148b expression, total RNA was polyadenylated using poly-A polymerase based First-Strand Synthesis kit (TaKaRa Bio, Japan) according to the manufacturer's protocol. .. To quantify WNT1 expression, total RNA (1ug) was converted to cDNA using M-MLV reverse transcriptase (Invitrogen).

Polymerase Chain Reaction:

Article Title: Complete Genome Sequence Analysis of Two Divergent Groups of Sweet potato chlorotic fleck virus Isolates Collected from Korea
Article Snippet: .. Reverse transcription polymerase chain reaction (RT-PCR) was performed as a two-step procedure; RT was conducted using Avian myeloblastosis virus (AMV) reverse transcriptase (Promega, USA), and PCR was carried out using high-fidelity LA taq polymerase (Takara, Japan). .. Specific PCR primer pairs were designed for primer walking and subsequent sequencing to obtain the complete genome sequences based on previously reported SPCFV nucleotide sequences ( , ).

Article Title: MiR-20a Promotes Cervical Cancer Proliferation and Metastasis In Vitro and In Vivo
Article Snippet: .. The reverse-transcription reactions were carried out using an MiraMasTM Kit (Bioo scientific, USA), which contains poly (A) polymerase used for polyadenylation of miRNA. qRT-PCR was performed using a standard SYBR Green PCR kit (Takara, Japan).The primers were synthesized (Shanghai GenePharma, China) as follows: miR-20a forwards primer: TAC GAT AAA GTG CTT ATA GTG CAG GTA G. U6 forwards primer: ATT GGA ACG ATA CAG AGA AGA TT. ..

Article Title: The pre- vertebrate origins of neurogenic placodes
Article Snippet: .. For the calcium imaging experiments, cDNA fragments containing the full-length coding regions of C. intestinalis CNGA, CNGB and CNGC (Ghost Gene IDs: KH.C2.249,KH.L42.6, and KH.C7.605, respectively) were amplified from a cDNA pool of mid tailbud embryos (CNGA) or larvae (CNGB and CNGC) by PCR using a thermostable DNA polymerase exhibiting proofreading activity (Takara LATaq; Takara Bio) with gene-specific primers (59-AGCTAACACAGATTTTAGTGTAATG-39 and 59-GTTGCGGGTAAATTACATGTC-39 for CNGA; 59-ATGGCATTGCACATAAATGCAA-39 and 59-AGCAAAGACTTTGGTAAACATCAG-39 for CNGB; 59-TAAGGTTGATACAGGTTTCATTGG-39 and 59-GACGTAATCATGACGAAACTGTG-39 for CNGC). .. The PCR products were subcloned into pcDNA6 plasmid vectors and sequenced on both strands using the cycle sequencing method with an Applied Biosystems 3100 Genetic Analyzer (Applied Biosystems, Foster City, CA, USA).

Article Title: RNAi in Cultured Drosophila Cells
Article Snippet: .. For PCR, substitute any thermostable DNA polymerase and reaction conditions with which you are familiar. .. We have found that the Clontech Advantage2 PCR enzyme/buffer mixture gives a high yield of product.

Article Title: Comparative Analysis of Gtf Isozyme Production and Diversity in Isolates of Streptococcus mutans with Different Biofilm Growth Phenotypes
Article Snippet: .. Amplification reactions were performed with a polymerase enzyme system specific for the generation of long PCR products (Advantage2; Clontech Labs, Inc.). ..

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    TaKaRa moloney murine leukemia virus superscript ii reverse transcriptase
    Moloney Murine Leukemia Virus Superscript Ii Reverse Transcriptase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/moloney murine leukemia virus superscript ii reverse transcriptase/product/TaKaRa
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    moloney murine leukemia virus superscript ii reverse transcriptase - by Bioz Stars, 2020-09
    85/100 stars
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