Review



rna decade marker  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier
Bioz Manufacturer Symbol Thermo Fisher manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 90
      Buy from Supplier

    Structured Review

    Thermo Fisher rna decade marker
    Rna Decade Marker, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rna+decade+marker/decade+marker+rna/us12123031-230-8-11
    Average 90 stars, based on 1 article reviews
    rna decade marker - by Bioz Stars, 2026-09
    90/100 stars

    Images

    Related Articles

    Marker:

    Article Title: RNA-RNA interactions between respiratory syncytial virus and miR-26 and miR-27 are associated with regulation of cell cycle and antiviral immunity.
    Article Snippet: Equal amounts of extracted RNA (3.5 μg) were mixed 1:1 with Gel Loading Buffer II (Invitrogen) and loaded on a 15% polyacrylamide / 8 M Urea gel. .. RNA Decade Marker (Thermo Scientific) was prepared according to the kit instructions, diluted in Gel Loading Buffer II, and run along the samples. .. The gel was run in 0.5x TBE at 120 V for 2 h. Samples were then transferred onto a HybondN membrane (Amersham Cat. RPN203N, 1 h at 80 V in 0.5 × TBE) and chemically crosslinked at 55 ◦C for 2 h using a solution of 0.16 M EDC ( N -(3-dimethylaminopropyl)N ′ -ethylcarbodiimide hydrochloride, Merck) with 0.13 M 1- methylimidazole in H 2 O (pH 8). ( 29 ) The membrane was washed once with 2 × SSC (20 × SSC, Fisher Scientific) and then pre-hybridised in 4 ml of Perfecthyb Plus hybridisation buffer (Sigma-Aldrich).

    Article Title: Regulation of CRISPR-associated rossman fold (CARF) domain containing proteins by oligoadenylates
    Article Snippet: .. The samples were analyzed in denaturing PAAG, with RNA Decade marker (Ambion) (M, first lane), RNA NSe alkaline hydrolysis marker (H, second lane) and RNA NSe partial RNase A (ThermoFisher) digest (A, third lane) for reference. ..

    Article Title: RNA–RNA interactions between respiratory syncytial virus and miR-26 and miR-27 are associated with regulation of cell cycle and antiviral immunity
    Article Snippet: Equal amounts of extracted RNA (3.5 μg) were mixed 1:1 with Gel Loading Buffer II (Invitrogen) and loaded on a 15% polyacrylamide/8 M Urea gel. .. RNA Decade Marker (Thermo Scientific) was prepared according to the kit instructions, diluted in Gel Loading Buffer II, and run along the samples. .. The gel was run in 0.5x TBE at 120 V for 2 h. Samples were then transferred onto a Hybond-N membrane (Amersham Cat. RPN203N, 1 h at 80 V in 0.5× TBE) and chemically crosslinked at 55°C for 2 h using a solution of 0.16 M EDC ( N -(3-dimethylaminopropyl)- N ′-ethylcarbodiimide hydrochloride, Merck) with 0.13 M 1- methylimidazole in H 2 O (pH 8). ( ) The membrane was washed once with 2× SSC (20× SSC, Fisher Scientific) and then pre-hybridised in 4 ml of Perfecthyb Plus hybridisation buffer (Sigma-Aldrich).

    Article Title: Dicer independent small RNAs associate with telomeric heterochromatin
    Article Snippet: .. For sizing control, the radio labeled RNA decade marker (Ambion) is shown on the left . ..

    Article Title: Nucleotide specificity of the human terminal nucleotidyltransferase Gld2 (TUT2)
    Article Snippet: .. The radiolabeled RNA Decade marker (Ambion) was used as reference. .. Twenty microliter reactions were performed containing 100 nM Gld2 in Buffer D and 2 μM of let-7a.

    Article Title: In silico prediction and expression profile analysis of small non-coding RNAs in Herbaspirillum seropedicae SmR1
    Article Snippet: Hybridizations were carried out overnight at 42 °C or 68 °C with the PerfectHyb Plus Hybridization Buffer (Sigma). .. RNA Decade marker (Ambion) was used when detecting non-coding RNAs up to 150 nt; for longer transcripts, the 100–1000 bp Ladder (Biotools) was used. ..

    Article Title: The appearance of pyrrolysine in tRNA His guanylyltransferase by neutral evolution
    Article Snippet: .. The RNA Decade marker (Ambion) was used. ..

    Article Title: Structural and functional basis of mammalian microRNA biogenesis by Dicer
    Article Snippet: The 5′-radiolabelled RNA was purified on G-25 columns (GE Healthcare) and diluted to a final concentration of 50 nM. .. The radiolabelled RNA Decade Marker (ThermoFisher Scientific) was prepared according to the manual. ..

    Control:

    Article Title: Dicer independent small RNAs associate with telomeric heterochromatin
    Article Snippet: .. For sizing control, the radio labeled RNA decade marker (Ambion) is shown on the left . ..

    Labeling:

    Article Title: Dicer independent small RNAs associate with telomeric heterochromatin
    Article Snippet: .. For sizing control, the radio labeled RNA decade marker (Ambion) is shown on the left . ..



    Similar Products

    decade marker rna  (New England Biolabs)
    99
    New England Biolabs decade marker rna
    Decade Marker Rna, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rna+decade+marker/Nuclease-free+Water/pmc04053197-47-8-37
    Average 99 stars, based on 1 article reviews
    decade marker rna - by Bioz Stars, 2026-09
    99/100 stars
    		  Buy from Supplier
    rna decade marker  (Thermo Fisher)
    90
    Thermo Fisher rna decade marker
    Rna Decade Marker, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rna+decade+marker/decade+marker+rna/us12123031-230-8-11
    Average 90 stars, based on 1 article reviews
    rna decade marker - by Bioz Stars, 2026-09
    90/100 stars
    		  Buy from Supplier
    century-plus decade rna markers  (Thermo Fisher)
    90
    Thermo Fisher century-plus decade rna markers
    Nascent transcripts of 21–23 nt, but not of 13–15 nt, are capped with fast-transcribing rL. ( A ) <t>N-RNA</t> was pre-incubated with rL and rP in the presence of ATP, CTP, and SAM. After adding saturating concentrations of UTP, GTP, and [α- 32 P]GDP, the reaction mixtures were incubated for the indicated periods. Cap-labeled transcripts were analyzed by 20% urea–PAGE followed by autoradiography. Marker RNAs are as follows: i, a mixture of 5′-capped oligo-RNAs with the VSV N mRNA start sequence (10 nt, Gppp N 10 ; 20 nt, Gppp N 20 ; 30 nt, Gppp N 30 ); ii, a hydroxide cleavage ladder of Gppp N 30 , 3′-dephosphorylated. The positions of the gel origin (ori.) and bottom (bot.) are indicated. ( B ) The transcripts of 21–23 nt synthesized in the presence of [α- 32 P]GDP for 60 s (see panel A, lane 4) were purified by urea–PAGE and digested with Nase P1. The digests were analyzed as in Figure . ( C ) Nascent transcripts were pulse-labeled with [α- 32 P]GDP for 60 sec as in panel A. After adding excess cold GDP, cap-labeled transcripts were chased for the indicated periods, and analyzed as in panel A. Maker lane iii <t>indicates</t> <t>Ambion</t> Decade Markers. ( D ) Cap-labeled nascent transcripts before and after chasing for 180 s (see panel C, lanes 1 and 4) were digested with Nase P1. The digests were analyzed as in Figure . ( E ) Nascent transcripts were synthesized in the presence of [α- 32 P]GDP (left scheme) or cold GDP (right scheme) for 60 s as in panel A. The latter RNAs were post-transcriptionally capped with [α- 32 P]GDP by rL. ( F ) Co-transcriptionally (lane 1) or post-transcriptionally (lane 2) capped transcripts were analyzed as in panel A. Marker RNAs are as follows: iv, a mixture of Gppp N 10 and Gppp N 20 ; v, a hydroxide cleavage ladder of Gppp N 20 , 3′-dephosphorylated.
    Century Plus Decade Rna Markers, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rna+decade+marker/pmc11381362-128-5-2
    Average 90 stars, based on 1 article reviews
    century-plus decade rna markers - by Bioz Stars, 2026-09
    90/100 stars
    		  Buy from Supplier
    decade rna marker  (Thermo Fisher)
    86
    Thermo Fisher decade rna marker
    Nascent transcripts of 21–23 nt, but not of 13–15 nt, are capped with fast-transcribing rL. ( A ) <t>N-RNA</t> was pre-incubated with rL and rP in the presence of ATP, CTP, and SAM. After adding saturating concentrations of UTP, GTP, and [α- 32 P]GDP, the reaction mixtures were incubated for the indicated periods. Cap-labeled transcripts were analyzed by 20% urea–PAGE followed by autoradiography. Marker RNAs are as follows: i, a mixture of 5′-capped oligo-RNAs with the VSV N mRNA start sequence (10 nt, Gppp N 10 ; 20 nt, Gppp N 20 ; 30 nt, Gppp N 30 ); ii, a hydroxide cleavage ladder of Gppp N 30 , 3′-dephosphorylated. The positions of the gel origin (ori.) and bottom (bot.) are indicated. ( B ) The transcripts of 21–23 nt synthesized in the presence of [α- 32 P]GDP for 60 s (see panel A, lane 4) were purified by urea–PAGE and digested with Nase P1. The digests were analyzed as in Figure . ( C ) Nascent transcripts were pulse-labeled with [α- 32 P]GDP for 60 sec as in panel A. After adding excess cold GDP, cap-labeled transcripts were chased for the indicated periods, and analyzed as in panel A. Maker lane iii <t>indicates</t> <t>Ambion</t> Decade Markers. ( D ) Cap-labeled nascent transcripts before and after chasing for 180 s (see panel C, lanes 1 and 4) were digested with Nase P1. The digests were analyzed as in Figure . ( E ) Nascent transcripts were synthesized in the presence of [α- 32 P]GDP (left scheme) or cold GDP (right scheme) for 60 s as in panel A. The latter RNAs were post-transcriptionally capped with [α- 32 P]GDP by rL. ( F ) Co-transcriptionally (lane 1) or post-transcriptionally (lane 2) capped transcripts were analyzed as in panel A. Marker RNAs are as follows: iv, a mixture of Gppp N 10 and Gppp N 20 ; v, a hydroxide cleavage ladder of Gppp N 20 , 3′-dephosphorylated.
    Decade Rna Marker, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rna+decade+marker/pm38375870-120-9-12
    Average 86 stars, based on 1 article reviews
    decade rna marker - by Bioz Stars, 2026-09
    86/100 stars
    		  Buy from Supplier
    rna decade™ marker  (Thermo Fisher)
    90
    Thermo Fisher rna decade™ marker
    Nascent transcripts of 21–23 nt, but not of 13–15 nt, are capped with fast-transcribing rL. ( A ) <t>N-RNA</t> was pre-incubated with rL and rP in the presence of ATP, CTP, and SAM. After adding saturating concentrations of UTP, GTP, and [α- 32 P]GDP, the reaction mixtures were incubated for the indicated periods. Cap-labeled transcripts were analyzed by 20% urea–PAGE followed by autoradiography. Marker RNAs are as follows: i, a mixture of 5′-capped oligo-RNAs with the VSV N mRNA start sequence (10 nt, Gppp N 10 ; 20 nt, Gppp N 20 ; 30 nt, Gppp N 30 ); ii, a hydroxide cleavage ladder of Gppp N 30 , 3′-dephosphorylated. The positions of the gel origin (ori.) and bottom (bot.) are indicated. ( B ) The transcripts of 21–23 nt synthesized in the presence of [α- 32 P]GDP for 60 s (see panel A, lane 4) were purified by urea–PAGE and digested with Nase P1. The digests were analyzed as in Figure . ( C ) Nascent transcripts were pulse-labeled with [α- 32 P]GDP for 60 sec as in panel A. After adding excess cold GDP, cap-labeled transcripts were chased for the indicated periods, and analyzed as in panel A. Maker lane iii <t>indicates</t> <t>Ambion</t> Decade Markers. ( D ) Cap-labeled nascent transcripts before and after chasing for 180 s (see panel C, lanes 1 and 4) were digested with Nase P1. The digests were analyzed as in Figure . ( E ) Nascent transcripts were synthesized in the presence of [α- 32 P]GDP (left scheme) or cold GDP (right scheme) for 60 s as in panel A. The latter RNAs were post-transcriptionally capped with [α- 32 P]GDP by rL. ( F ) Co-transcriptionally (lane 1) or post-transcriptionally (lane 2) capped transcripts were analyzed as in panel A. Marker RNAs are as follows: iv, a mixture of Gppp N 10 and Gppp N 20 ; v, a hydroxide cleavage ladder of Gppp N 20 , 3′-dephosphorylated.
    Rna Decade™ Marker, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rna+decade+marker/pmc03132168-367-17-20
    Average 90 stars, based on 1 article reviews
    rna decade™ marker - by Bioz Stars, 2026-09
    90/100 stars
    		  Buy from Supplier
    decade™ marker rna  (Thermo Fisher)
    90
    Thermo Fisher decade™ marker rna
    Nascent transcripts of 21–23 nt, but not of 13–15 nt, are capped with fast-transcribing rL. ( A ) <t>N-RNA</t> was pre-incubated with rL and rP in the presence of ATP, CTP, and SAM. After adding saturating concentrations of UTP, GTP, and [α- 32 P]GDP, the reaction mixtures were incubated for the indicated periods. Cap-labeled transcripts were analyzed by 20% urea–PAGE followed by autoradiography. Marker RNAs are as follows: i, a mixture of 5′-capped oligo-RNAs with the VSV N mRNA start sequence (10 nt, Gppp N 10 ; 20 nt, Gppp N 20 ; 30 nt, Gppp N 30 ); ii, a hydroxide cleavage ladder of Gppp N 30 , 3′-dephosphorylated. The positions of the gel origin (ori.) and bottom (bot.) are indicated. ( B ) The transcripts of 21–23 nt synthesized in the presence of [α- 32 P]GDP for 60 s (see panel A, lane 4) were purified by urea–PAGE and digested with Nase P1. The digests were analyzed as in Figure . ( C ) Nascent transcripts were pulse-labeled with [α- 32 P]GDP for 60 sec as in panel A. After adding excess cold GDP, cap-labeled transcripts were chased for the indicated periods, and analyzed as in panel A. Maker lane iii <t>indicates</t> <t>Ambion</t> Decade Markers. ( D ) Cap-labeled nascent transcripts before and after chasing for 180 s (see panel C, lanes 1 and 4) were digested with Nase P1. The digests were analyzed as in Figure . ( E ) Nascent transcripts were synthesized in the presence of [α- 32 P]GDP (left scheme) or cold GDP (right scheme) for 60 s as in panel A. The latter RNAs were post-transcriptionally capped with [α- 32 P]GDP by rL. ( F ) Co-transcriptionally (lane 1) or post-transcriptionally (lane 2) capped transcripts were analyzed as in panel A. Marker RNAs are as follows: iv, a mixture of Gppp N 10 and Gppp N 20 ; v, a hydroxide cleavage ladder of Gppp N 20 , 3′-dephosphorylated.
    Decade™ Marker Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rna+decade+marker/pmc06152437-230-1-4
    Average 90 stars, based on 1 article reviews
    decade™ marker rna - by Bioz Stars, 2026-09
    90/100 stars
    		  Buy from Supplier
    decade marker rna (100 ng)  (Thermo Fisher)
    90
    Thermo Fisher decade marker rna (100 ng)
    Nascent transcripts of 21–23 nt, but not of 13–15 nt, are capped with fast-transcribing rL. ( A ) <t>N-RNA</t> was pre-incubated with rL and rP in the presence of ATP, CTP, and SAM. After adding saturating concentrations of UTP, GTP, and [α- 32 P]GDP, the reaction mixtures were incubated for the indicated periods. Cap-labeled transcripts were analyzed by 20% urea–PAGE followed by autoradiography. Marker RNAs are as follows: i, a mixture of 5′-capped oligo-RNAs with the VSV N mRNA start sequence (10 nt, Gppp N 10 ; 20 nt, Gppp N 20 ; 30 nt, Gppp N 30 ); ii, a hydroxide cleavage ladder of Gppp N 30 , 3′-dephosphorylated. The positions of the gel origin (ori.) and bottom (bot.) are indicated. ( B ) The transcripts of 21–23 nt synthesized in the presence of [α- 32 P]GDP for 60 s (see panel A, lane 4) were purified by urea–PAGE and digested with Nase P1. The digests were analyzed as in Figure . ( C ) Nascent transcripts were pulse-labeled with [α- 32 P]GDP for 60 sec as in panel A. After adding excess cold GDP, cap-labeled transcripts were chased for the indicated periods, and analyzed as in panel A. Maker lane iii <t>indicates</t> <t>Ambion</t> Decade Markers. ( D ) Cap-labeled nascent transcripts before and after chasing for 180 s (see panel C, lanes 1 and 4) were digested with Nase P1. The digests were analyzed as in Figure . ( E ) Nascent transcripts were synthesized in the presence of [α- 32 P]GDP (left scheme) or cold GDP (right scheme) for 60 s as in panel A. The latter RNAs were post-transcriptionally capped with [α- 32 P]GDP by rL. ( F ) Co-transcriptionally (lane 1) or post-transcriptionally (lane 2) capped transcripts were analyzed as in panel A. Marker RNAs are as follows: iv, a mixture of Gppp N 10 and Gppp N 20 ; v, a hydroxide cleavage ladder of Gppp N 20 , 3′-dephosphorylated.
    Decade Marker Rna (100 Ng), supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rna+decade+marker/pmc04053197-47-8-3
    Average 90 stars, based on 1 article reviews
    decade marker rna (100 ng) - by Bioz Stars, 2026-09
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    Nascent transcripts of 21–23 nt, but not of 13–15 nt, are capped with fast-transcribing rL. ( A ) N-RNA was pre-incubated with rL and rP in the presence of ATP, CTP, and SAM. After adding saturating concentrations of UTP, GTP, and [α- 32 P]GDP, the reaction mixtures were incubated for the indicated periods. Cap-labeled transcripts were analyzed by 20% urea–PAGE followed by autoradiography. Marker RNAs are as follows: i, a mixture of 5′-capped oligo-RNAs with the VSV N mRNA start sequence (10 nt, Gppp N 10 ; 20 nt, Gppp N 20 ; 30 nt, Gppp N 30 ); ii, a hydroxide cleavage ladder of Gppp N 30 , 3′-dephosphorylated. The positions of the gel origin (ori.) and bottom (bot.) are indicated. ( B ) The transcripts of 21–23 nt synthesized in the presence of [α- 32 P]GDP for 60 s (see panel A, lane 4) were purified by urea–PAGE and digested with Nase P1. The digests were analyzed as in Figure . ( C ) Nascent transcripts were pulse-labeled with [α- 32 P]GDP for 60 sec as in panel A. After adding excess cold GDP, cap-labeled transcripts were chased for the indicated periods, and analyzed as in panel A. Maker lane iii indicates Ambion Decade Markers. ( D ) Cap-labeled nascent transcripts before and after chasing for 180 s (see panel C, lanes 1 and 4) were digested with Nase P1. The digests were analyzed as in Figure . ( E ) Nascent transcripts were synthesized in the presence of [α- 32 P]GDP (left scheme) or cold GDP (right scheme) for 60 s as in panel A. The latter RNAs were post-transcriptionally capped with [α- 32 P]GDP by rL. ( F ) Co-transcriptionally (lane 1) or post-transcriptionally (lane 2) capped transcripts were analyzed as in panel A. Marker RNAs are as follows: iv, a mixture of Gppp N 10 and Gppp N 20 ; v, a hydroxide cleavage ladder of Gppp N 20 , 3′-dephosphorylated.

    Journal: Nucleic Acids Research

    Article Title: The complete pathway for co-transcriptional mRNA maturation within a large protein of a non-segmented negative-strand RNA virus

    doi: 10.1093/nar/gkae659

    Figure Lengend Snippet: Nascent transcripts of 21–23 nt, but not of 13–15 nt, are capped with fast-transcribing rL. ( A ) N-RNA was pre-incubated with rL and rP in the presence of ATP, CTP, and SAM. After adding saturating concentrations of UTP, GTP, and [α- 32 P]GDP, the reaction mixtures were incubated for the indicated periods. Cap-labeled transcripts were analyzed by 20% urea–PAGE followed by autoradiography. Marker RNAs are as follows: i, a mixture of 5′-capped oligo-RNAs with the VSV N mRNA start sequence (10 nt, Gppp N 10 ; 20 nt, Gppp N 20 ; 30 nt, Gppp N 30 ); ii, a hydroxide cleavage ladder of Gppp N 30 , 3′-dephosphorylated. The positions of the gel origin (ori.) and bottom (bot.) are indicated. ( B ) The transcripts of 21–23 nt synthesized in the presence of [α- 32 P]GDP for 60 s (see panel A, lane 4) were purified by urea–PAGE and digested with Nase P1. The digests were analyzed as in Figure . ( C ) Nascent transcripts were pulse-labeled with [α- 32 P]GDP for 60 sec as in panel A. After adding excess cold GDP, cap-labeled transcripts were chased for the indicated periods, and analyzed as in panel A. Maker lane iii indicates Ambion Decade Markers. ( D ) Cap-labeled nascent transcripts before and after chasing for 180 s (see panel C, lanes 1 and 4) were digested with Nase P1. The digests were analyzed as in Figure . ( E ) Nascent transcripts were synthesized in the presence of [α- 32 P]GDP (left scheme) or cold GDP (right scheme) for 60 s as in panel A. The latter RNAs were post-transcriptionally capped with [α- 32 P]GDP by rL. ( F ) Co-transcriptionally (lane 1) or post-transcriptionally (lane 2) capped transcripts were analyzed as in panel A. Marker RNAs are as follows: iv, a mixture of Gppp N 10 and Gppp N 20 ; v, a hydroxide cleavage ladder of Gppp N 20 , 3′-dephosphorylated.

    Article Snippet: 32 P-Labeled Ambion Century-Plus and Decade RNA markers (Life Technologies/Thermo Fisher Scientific) were prepared according to the manufacturer's protocols.

    Techniques: Incubation, Labeling, Autoradiography, Marker, Sequencing, Synthesized, Purification

    L methylates the cap on pre-mRNAs during mRNA chain elongation. ( A ) Diagrams of each pulse-chase experiment are shown. ( B and C ) After preincubation of N-RNA, rL, and rP in the presence of ATP, CTP, and SAM, nascent transcripts were pulse-labeled with [α- 32 P]GDP in the presence of the limiting concentration of GTP. After adding excess concentrations of cold GDP and GTP, cap-labeled nascent transcripts were chased for the indicated periods (10 sec to 11 min) (see panel A, middle) and analyzed by 20% (B) or 5% (C) urea–PAGE followed by autoradiography. An asterisk indicates an abortive transcript of 40 nt. Marker RNAs are as follows: viii, Ambion Century-Plus RNA Markers, ix, deadenylated VSV N , P / M , and G mRNAs. ( D ) Cap-labeled transcripts chased for 11 min (lane 1) were digested with RNase H in the presence of oligo(dT) (lane 2) and analyzed as in panel C. ( E ) Cap-labeled transcripts chased for the indicated periods were digested with Nase P1. The digests were analyzed as in Figure . ( F ) Pulse-chase reactions performed in the absence of SAM (see panel A, lower). During the chase period, the reactions were received water or SAM (final concentration: 30 μM) at the indicated time point and further incubated for 3 min. The cap methylation status was analyzed as in panel E. ( G – I ) Chase reactions were terminated at the indicated time point and subjected to ultracentrifugation to separate free transcripts (Supernatants, Sup) from RNP-associated transcripts (Precipitates, Ppt). Transcripts in each fraction were analyzed by 20% (H) or 5% (I) urea–PAGE followed by autoradiography.

    Journal: Nucleic Acids Research

    Article Title: The complete pathway for co-transcriptional mRNA maturation within a large protein of a non-segmented negative-strand RNA virus

    doi: 10.1093/nar/gkae659

    Figure Lengend Snippet: L methylates the cap on pre-mRNAs during mRNA chain elongation. ( A ) Diagrams of each pulse-chase experiment are shown. ( B and C ) After preincubation of N-RNA, rL, and rP in the presence of ATP, CTP, and SAM, nascent transcripts were pulse-labeled with [α- 32 P]GDP in the presence of the limiting concentration of GTP. After adding excess concentrations of cold GDP and GTP, cap-labeled nascent transcripts were chased for the indicated periods (10 sec to 11 min) (see panel A, middle) and analyzed by 20% (B) or 5% (C) urea–PAGE followed by autoradiography. An asterisk indicates an abortive transcript of 40 nt. Marker RNAs are as follows: viii, Ambion Century-Plus RNA Markers, ix, deadenylated VSV N , P / M , and G mRNAs. ( D ) Cap-labeled transcripts chased for 11 min (lane 1) were digested with RNase H in the presence of oligo(dT) (lane 2) and analyzed as in panel C. ( E ) Cap-labeled transcripts chased for the indicated periods were digested with Nase P1. The digests were analyzed as in Figure . ( F ) Pulse-chase reactions performed in the absence of SAM (see panel A, lower). During the chase period, the reactions were received water or SAM (final concentration: 30 μM) at the indicated time point and further incubated for 3 min. The cap methylation status was analyzed as in panel E. ( G – I ) Chase reactions were terminated at the indicated time point and subjected to ultracentrifugation to separate free transcripts (Supernatants, Sup) from RNP-associated transcripts (Precipitates, Ppt). Transcripts in each fraction were analyzed by 20% (H) or 5% (I) urea–PAGE followed by autoradiography.

    Article Snippet: 32 P-Labeled Ambion Century-Plus and Decade RNA markers (Life Technologies/Thermo Fisher Scientific) were prepared according to the manufacturer's protocols.

    Techniques: Pulse Chase, Labeling, Concentration Assay, Autoradiography, Marker, Incubation, Methylation