Structured Review

Promega oligo dt
The 2b-block SSO inhibits glioblastoma formation in vivo . ( A ) Schematic representation of injection of cells into the murine brain. ( B ) mCherry-labeled U87MG cells resuspended in PBS containing 2′-OMe 2b-block SSO or SCR <t>oligo</t> 12.5 μg/μl were injected intracranially into the striatum on both sides. After 14 days the mice were sacrificed and brains were photographed under a fluorescent dissecting microscope. ( C ) RT-PCR analysis of <t>RNA</t> isolated from the tumors described in (B). * P = 0.0087 two-sided, n = 7 for each group.
Oligo Dt, supplied by Promega, used in various techniques. Bioz Stars score: 94/100, based on 396 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 94 stars, based on 396 article reviews
Price from $9.99 to $1999.99
oligo dt - by Bioz Stars, 2020-08
94/100 stars

Images

1) Product Images from "Modulation of MKNK2 alternative splicing by splice-switching oligonucleotides as a novel approach for glioblastoma treatment"

Article Title: Modulation of MKNK2 alternative splicing by splice-switching oligonucleotides as a novel approach for glioblastoma treatment

Journal: Nucleic Acids Research

doi: 10.1093/nar/gky921

The 2b-block SSO inhibits glioblastoma formation in vivo . ( A ) Schematic representation of injection of cells into the murine brain. ( B ) mCherry-labeled U87MG cells resuspended in PBS containing 2′-OMe 2b-block SSO or SCR oligo 12.5 μg/μl were injected intracranially into the striatum on both sides. After 14 days the mice were sacrificed and brains were photographed under a fluorescent dissecting microscope. ( C ) RT-PCR analysis of RNA isolated from the tumors described in (B). * P = 0.0087 two-sided, n = 7 for each group.
Figure Legend Snippet: The 2b-block SSO inhibits glioblastoma formation in vivo . ( A ) Schematic representation of injection of cells into the murine brain. ( B ) mCherry-labeled U87MG cells resuspended in PBS containing 2′-OMe 2b-block SSO or SCR oligo 12.5 μg/μl were injected intracranially into the striatum on both sides. After 14 days the mice were sacrificed and brains were photographed under a fluorescent dissecting microscope. ( C ) RT-PCR analysis of RNA isolated from the tumors described in (B). * P = 0.0087 two-sided, n = 7 for each group.

Techniques Used: Blocking Assay, In Vivo, Injection, Labeling, Mouse Assay, Microscopy, Reverse Transcription Polymerase Chain Reaction, Isolation

2) Product Images from "Promiscuous recombination of LoxP alleles during gametogenesis in cornea Cre driver mice"

Article Title: Promiscuous recombination of LoxP alleles during gametogenesis in cornea Cre driver mice

Journal: Molecular Vision

doi:

Reverse transcription polymerase chain reaction for detection of keratocan, keratin 12 and Cre mRNAs in testis. Total RNAs were isolated from four pooled corneas and individual testis of two Kera-Cre mice with TRIzol Reagent (Invitrogen), and then the total RNAs were dissolved in DEPC-Water and stored at −80 °C until use. Ten micrograms (testis) and 3.5 μg of total RNA were annealed to oligo-dT and reverse transcribed with kits from Promega according to the manufacturer’s instruction. Single stranded cDNA was subjected to PCR reactions using primer pairs for keratocan ( Kera ), keratin 12 ( K12 ), Cre recombinase ( Cre ), and glyceraldehydes phosphate dehydrogenase ( GAPDH ) as described in Methods. A : Use of primers in exons 1 and 3 of Krt12 generates a 408 bp RT–PCR product with total RNA from the cornea, but not that from testis; B : use of primers in exons 7 and 8 of Krt12 , a 410 bp RT–PCR product was detected with RNA from both corneas and testis; C : use of primers in exons 2 and 3 of Kera produces a 350 bp RT–PCR product with RNA from cornea and testis; D , Use primer of exon 1 of Kera and primer of IRES yields a 480 bp RT–PCR product in RNA from both cornea and testis of Kera-Cre mice; E : primers of exon 6 and 7 of GAPDH gene, a 200 bp transcript was used as a positive control RT–PCR with RNA from both cornea and testis. Lane 1, testis 1; lane 2, testis 2; lane 3, cornea; lane 4, control (no cDNA added); M, 1 kb DNA markers.
Figure Legend Snippet: Reverse transcription polymerase chain reaction for detection of keratocan, keratin 12 and Cre mRNAs in testis. Total RNAs were isolated from four pooled corneas and individual testis of two Kera-Cre mice with TRIzol Reagent (Invitrogen), and then the total RNAs were dissolved in DEPC-Water and stored at −80 °C until use. Ten micrograms (testis) and 3.5 μg of total RNA were annealed to oligo-dT and reverse transcribed with kits from Promega according to the manufacturer’s instruction. Single stranded cDNA was subjected to PCR reactions using primer pairs for keratocan ( Kera ), keratin 12 ( K12 ), Cre recombinase ( Cre ), and glyceraldehydes phosphate dehydrogenase ( GAPDH ) as described in Methods. A : Use of primers in exons 1 and 3 of Krt12 generates a 408 bp RT–PCR product with total RNA from the cornea, but not that from testis; B : use of primers in exons 7 and 8 of Krt12 , a 410 bp RT–PCR product was detected with RNA from both corneas and testis; C : use of primers in exons 2 and 3 of Kera produces a 350 bp RT–PCR product with RNA from cornea and testis; D , Use primer of exon 1 of Kera and primer of IRES yields a 480 bp RT–PCR product in RNA from both cornea and testis of Kera-Cre mice; E : primers of exon 6 and 7 of GAPDH gene, a 200 bp transcript was used as a positive control RT–PCR with RNA from both cornea and testis. Lane 1, testis 1; lane 2, testis 2; lane 3, cornea; lane 4, control (no cDNA added); M, 1 kb DNA markers.

Techniques Used: Reverse Transcription Polymerase Chain Reaction, Isolation, Mouse Assay, Polymerase Chain Reaction, Positive Control

Related Articles

SYBR Green Assay:

Article Title: Neutral sphingomyelinase 2 modulates cytotoxic effects of protopanaxadiol on different human cancer cells
Article Snippet: .. Total RNA was isolated from cultured cells using Trizol (Invitrogen). cDNAs were synthesized using oligo (dT) and SuperScript III reverse transcriptase (Promega, Madison, WI, USA), then amplified using iQ SYBR Green PCR Master Mix (Bio-Rad Laboratories, Hercules, CA, USA) in conjunction with Mx3005P QPCR Systems (Agilent Technologies, Santa Clara, CA, USA). .. Primer sets were as follows: nSMase 2 qPCR primer P109289 (Bioneer); glyceraldehyde-3-phosphate dehydrogenase, 5′-TGGTATCGTGGAAGGACTCATGAC-3′ (forward) and 5′-ATGCCAGTGAGCTTCCCGTTCAGC -3′ (reverse).

Amplification:

Article Title: Neutral sphingomyelinase 2 modulates cytotoxic effects of protopanaxadiol on different human cancer cells
Article Snippet: .. Total RNA was isolated from cultured cells using Trizol (Invitrogen). cDNAs were synthesized using oligo (dT) and SuperScript III reverse transcriptase (Promega, Madison, WI, USA), then amplified using iQ SYBR Green PCR Master Mix (Bio-Rad Laboratories, Hercules, CA, USA) in conjunction with Mx3005P QPCR Systems (Agilent Technologies, Santa Clara, CA, USA). .. Primer sets were as follows: nSMase 2 qPCR primer P109289 (Bioneer); glyceraldehyde-3-phosphate dehydrogenase, 5′-TGGTATCGTGGAAGGACTCATGAC-3′ (forward) and 5′-ATGCCAGTGAGCTTCCCGTTCAGC -3′ (reverse).

Synthesized:

Article Title: Modulation of MKNK2 alternative splicing by splice-switching oligonucleotides as a novel approach for glioblastoma treatment
Article Snippet: .. Total RNA was isolated and cDNA was synthesized from 1 μg RNA using MMLV reverse transcriptase and oligo dT in a final volume of 25 μl according to the manufacturer's instructions (Promega). .. Polymerase chain reacion (PCR) was conducted on 1 μl of cDNA by KAPA 2G Fast HS ReadyMix PCR kit (KAPA Biosystems).

Article Title: Neutral sphingomyelinase 2 modulates cytotoxic effects of protopanaxadiol on different human cancer cells
Article Snippet: .. Total RNA was isolated from cultured cells using Trizol (Invitrogen). cDNAs were synthesized using oligo (dT) and SuperScript III reverse transcriptase (Promega, Madison, WI, USA), then amplified using iQ SYBR Green PCR Master Mix (Bio-Rad Laboratories, Hercules, CA, USA) in conjunction with Mx3005P QPCR Systems (Agilent Technologies, Santa Clara, CA, USA). .. Primer sets were as follows: nSMase 2 qPCR primer P109289 (Bioneer); glyceraldehyde-3-phosphate dehydrogenase, 5′-TGGTATCGTGGAAGGACTCATGAC-3′ (forward) and 5′-ATGCCAGTGAGCTTCCCGTTCAGC -3′ (reverse).

Isolation:

Article Title: Modulation of MKNK2 alternative splicing by splice-switching oligonucleotides as a novel approach for glioblastoma treatment
Article Snippet: .. Total RNA was isolated and cDNA was synthesized from 1 μg RNA using MMLV reverse transcriptase and oligo dT in a final volume of 25 μl according to the manufacturer's instructions (Promega). .. Polymerase chain reacion (PCR) was conducted on 1 μl of cDNA by KAPA 2G Fast HS ReadyMix PCR kit (KAPA Biosystems).

Article Title: Neutral sphingomyelinase 2 modulates cytotoxic effects of protopanaxadiol on different human cancer cells
Article Snippet: .. Total RNA was isolated from cultured cells using Trizol (Invitrogen). cDNAs were synthesized using oligo (dT) and SuperScript III reverse transcriptase (Promega, Madison, WI, USA), then amplified using iQ SYBR Green PCR Master Mix (Bio-Rad Laboratories, Hercules, CA, USA) in conjunction with Mx3005P QPCR Systems (Agilent Technologies, Santa Clara, CA, USA). .. Primer sets were as follows: nSMase 2 qPCR primer P109289 (Bioneer); glyceraldehyde-3-phosphate dehydrogenase, 5′-TGGTATCGTGGAAGGACTCATGAC-3′ (forward) and 5′-ATGCCAGTGAGCTTCCCGTTCAGC -3′ (reverse).

Random Hexamer Labeling:

Article Title: Curcumin Blocks Interleukin-1 Signaling in Chondrosarcoma Cells
Article Snippet: .. 1 µg of total RNA was reverse transcribed using BioScript Moloney Murine Leukaemia Virus (MMLV) Reverse Transcriptase (BioLine, Luckenwalde, Germany) with oligo dT and random hexamer primers (Promega, Madison, WI, USA) in separate reactions. .. Quantitative PCR was performed using the LC Fast Start DNA Master Plus Sybr Green PCR Mix and a LightCycler 2.0 (Roche, Mannheim, Germany).

Cell Culture:

Article Title: Neutral sphingomyelinase 2 modulates cytotoxic effects of protopanaxadiol on different human cancer cells
Article Snippet: .. Total RNA was isolated from cultured cells using Trizol (Invitrogen). cDNAs were synthesized using oligo (dT) and SuperScript III reverse transcriptase (Promega, Madison, WI, USA), then amplified using iQ SYBR Green PCR Master Mix (Bio-Rad Laboratories, Hercules, CA, USA) in conjunction with Mx3005P QPCR Systems (Agilent Technologies, Santa Clara, CA, USA). .. Primer sets were as follows: nSMase 2 qPCR primer P109289 (Bioneer); glyceraldehyde-3-phosphate dehydrogenase, 5′-TGGTATCGTGGAAGGACTCATGAC-3′ (forward) and 5′-ATGCCAGTGAGCTTCCCGTTCAGC -3′ (reverse).

Real-time Polymerase Chain Reaction:

Article Title: Neutral sphingomyelinase 2 modulates cytotoxic effects of protopanaxadiol on different human cancer cells
Article Snippet: .. Total RNA was isolated from cultured cells using Trizol (Invitrogen). cDNAs were synthesized using oligo (dT) and SuperScript III reverse transcriptase (Promega, Madison, WI, USA), then amplified using iQ SYBR Green PCR Master Mix (Bio-Rad Laboratories, Hercules, CA, USA) in conjunction with Mx3005P QPCR Systems (Agilent Technologies, Santa Clara, CA, USA). .. Primer sets were as follows: nSMase 2 qPCR primer P109289 (Bioneer); glyceraldehyde-3-phosphate dehydrogenase, 5′-TGGTATCGTGGAAGGACTCATGAC-3′ (forward) and 5′-ATGCCAGTGAGCTTCCCGTTCAGC -3′ (reverse).

Incubation:

Article Title: Stage-Regulated GFP Expression in Trypanosoma cruzi: Applications from Host-Parasite Interactions to Drug Screening
Article Snippet: .. The first-strand cDNA was obtained by mixing 1 µg of total RNA and 1 µM oligo dT and incubating at 70°C for 10 min. We then mixed 4 µl of Improm-II buffer (Promega, Madison, USA), 3 mM MgCl2 , 0.5 mM of each dNTP, 40 U RNaseOUT (Invitrogen) and 2 µl Improm-II reverse transcriptase (Promega) in a final volume of 20 µl and incubated the mixture for 2 h at 42°C. .. The product was then purified with Microcon(r) YM-30 (Millipore, Massachusetts, USA) and resuspended at a concentration of 2 ng µl−1 in water.

Polymerase Chain Reaction:

Article Title: Neutral sphingomyelinase 2 modulates cytotoxic effects of protopanaxadiol on different human cancer cells
Article Snippet: .. Total RNA was isolated from cultured cells using Trizol (Invitrogen). cDNAs were synthesized using oligo (dT) and SuperScript III reverse transcriptase (Promega, Madison, WI, USA), then amplified using iQ SYBR Green PCR Master Mix (Bio-Rad Laboratories, Hercules, CA, USA) in conjunction with Mx3005P QPCR Systems (Agilent Technologies, Santa Clara, CA, USA). .. Primer sets were as follows: nSMase 2 qPCR primer P109289 (Bioneer); glyceraldehyde-3-phosphate dehydrogenase, 5′-TGGTATCGTGGAAGGACTCATGAC-3′ (forward) and 5′-ATGCCAGTGAGCTTCCCGTTCAGC -3′ (reverse).

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    Promega oligo dt primer
    USP21 modulates Hh signaling and Gli1 transcriptional activity. (A) NIH3T3 cells were treated with USP21-targeting siRNA oligonucleotides (#10, #11, #12) or a non-targeting control (NT3), serum-starved and then incubated with SAG (100 nM) for 4 h before mRNA extraction. USP21 and Gli1 mRNA levels were measured by performing quantitative reverse-transcription PCR and normalised to <t>RNA</t> levels of Pol2 ( n =3 independent experiments, mean±s.d.). x -axis labels given in bottom panel also apply to the top panel. (B) HEK293T cells were treated with USP21-targeting siRNA oligonucleotides (#5, #6) or non-targeting control <t>oligo</t> (NT3), or were treated with transfection reagent alone (Mock). 24 h before lysis, cells were transfected with Gli-responsive firefly luciferase and Renilla luciferase reporters (pGLB3B-12Gli-Luc and pRL-Renilla-TK), as well as with a Gli1 expression construct (HA–Gli1). Efficiency of the knockdown was assessed by western blotting ( n =2, bars show the range). (C) HEK293T cells were transfected with Gli-responsive firefly luciferase, Renilla luciferase reporters, HA–Gli1 and the indicated amounts of GFP, GFP–USP21 or GFP–USP21CS plasmids. Protein expression was assessed by western blotting (IB, immunoblot) for GFP. (GFP, n =5; USP21 and USP21CS, n =6; mean±s.d.; one-way ANOVA and Dunnett's test, ** P
    Oligo Dt Primer, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 317 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/oligo dt primer/product/Promega
    Average 99 stars, based on 317 article reviews
    Price from $9.99 to $1999.99
    oligo dt primer - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    92
    Promega oligo dt chromatography
    : localisation of <t>mRNA.</t> (A) Fluorescent in situ hybridisation assay using specific <t>oligo</t> (dT) probe. As the control, RNase A was incubated with the parasites before probe hybridisation. DAPI: DNA stained with DAPI; mRNA: mRNA detection; granules: labelling of pixels with high intensity fluorescence signals; N: nucleus; K: kinetoplast. Scale bar = 5 µm. (B) Histogram of mRNA granule counts per cell.
    Oligo Dt Chromatography, supplied by Promega, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/oligo dt chromatography/product/Promega
    Average 92 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    oligo dt chromatography - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    Image Search Results


    USP21 modulates Hh signaling and Gli1 transcriptional activity. (A) NIH3T3 cells were treated with USP21-targeting siRNA oligonucleotides (#10, #11, #12) or a non-targeting control (NT3), serum-starved and then incubated with SAG (100 nM) for 4 h before mRNA extraction. USP21 and Gli1 mRNA levels were measured by performing quantitative reverse-transcription PCR and normalised to RNA levels of Pol2 ( n =3 independent experiments, mean±s.d.). x -axis labels given in bottom panel also apply to the top panel. (B) HEK293T cells were treated with USP21-targeting siRNA oligonucleotides (#5, #6) or non-targeting control oligo (NT3), or were treated with transfection reagent alone (Mock). 24 h before lysis, cells were transfected with Gli-responsive firefly luciferase and Renilla luciferase reporters (pGLB3B-12Gli-Luc and pRL-Renilla-TK), as well as with a Gli1 expression construct (HA–Gli1). Efficiency of the knockdown was assessed by western blotting ( n =2, bars show the range). (C) HEK293T cells were transfected with Gli-responsive firefly luciferase, Renilla luciferase reporters, HA–Gli1 and the indicated amounts of GFP, GFP–USP21 or GFP–USP21CS plasmids. Protein expression was assessed by western blotting (IB, immunoblot) for GFP. (GFP, n =5; USP21 and USP21CS, n =6; mean±s.d.; one-way ANOVA and Dunnett's test, ** P

    Journal: Journal of Cell Science

    Article Title: The centrosomal deubiquitylase USP21 regulates Gli1 transcriptional activity and stability

    doi: 10.1242/jcs.188516

    Figure Lengend Snippet: USP21 modulates Hh signaling and Gli1 transcriptional activity. (A) NIH3T3 cells were treated with USP21-targeting siRNA oligonucleotides (#10, #11, #12) or a non-targeting control (NT3), serum-starved and then incubated with SAG (100 nM) for 4 h before mRNA extraction. USP21 and Gli1 mRNA levels were measured by performing quantitative reverse-transcription PCR and normalised to RNA levels of Pol2 ( n =3 independent experiments, mean±s.d.). x -axis labels given in bottom panel also apply to the top panel. (B) HEK293T cells were treated with USP21-targeting siRNA oligonucleotides (#5, #6) or non-targeting control oligo (NT3), or were treated with transfection reagent alone (Mock). 24 h before lysis, cells were transfected with Gli-responsive firefly luciferase and Renilla luciferase reporters (pGLB3B-12Gli-Luc and pRL-Renilla-TK), as well as with a Gli1 expression construct (HA–Gli1). Efficiency of the knockdown was assessed by western blotting ( n =2, bars show the range). (C) HEK293T cells were transfected with Gli-responsive firefly luciferase, Renilla luciferase reporters, HA–Gli1 and the indicated amounts of GFP, GFP–USP21 or GFP–USP21CS plasmids. Protein expression was assessed by western blotting (IB, immunoblot) for GFP. (GFP, n =5; USP21 and USP21CS, n =6; mean±s.d.; one-way ANOVA and Dunnett's test, ** P

    Article Snippet: Quantitative reverse-transcription PCR Cells were lysed, and mRNA was extracted using the RNAeasy mini kit (Qiagen). cDNA synthesis was performed using 1 µg RNA with RevertAid H-minus M-MuLV reverse transcriptase (Fermentas) using an oligo-dT primer (Promega).

    Techniques: Activity Assay, Incubation, Polymerase Chain Reaction, Transfection, Lysis, Luciferase, Expressing, Construct, Western Blot

    FcRn expression in normal adult human small intestine and human intestinal epithelial cell lines. Western blots of total cellular protein (13 μg protein per lane, a ; 10 μg protein per lane, b ) isolated from the indicated source using affinity-purified rabbit antisera raised against amino acids 112–125 ( a ) or amino acids 174–188 ( b ). ( c ) RT-PCR detection of FcγRI transcripts. Total RNA (2 μg) from T84 (lanes 3 and 4), MOLT-4 (lanes 5 and 6; negative control), and U937 (lanes 1 and 2; positive control) cell lines was incubated with an oligo-dT primer with (odd-numbered lanes) or without (even-numbered lanes) avian myeloblastosis virus–RT (AMV-RT), and a nested PCR was performed with primers specific for FcγRI cDNA (top) or for β-actin (bottom).

    Journal: Journal of Clinical Investigation

    Article Title: Bidirectional FcRn-dependent IgG transport in a polarized human intestinal epithelial cell line

    doi:

    Figure Lengend Snippet: FcRn expression in normal adult human small intestine and human intestinal epithelial cell lines. Western blots of total cellular protein (13 μg protein per lane, a ; 10 μg protein per lane, b ) isolated from the indicated source using affinity-purified rabbit antisera raised against amino acids 112–125 ( a ) or amino acids 174–188 ( b ). ( c ) RT-PCR detection of FcγRI transcripts. Total RNA (2 μg) from T84 (lanes 3 and 4), MOLT-4 (lanes 5 and 6; negative control), and U937 (lanes 1 and 2; positive control) cell lines was incubated with an oligo-dT primer with (odd-numbered lanes) or without (even-numbered lanes) avian myeloblastosis virus–RT (AMV-RT), and a nested PCR was performed with primers specific for FcγRI cDNA (top) or for β-actin (bottom).

    Article Snippet: RNA (2 μg) was reverse-transcribed to cDNA with an oligo-dT primer (Promega Corp., Madison, Wisconsin, USA) and avian myeloblastosis virus reverse transcriptase (Promega Corp.).

    Techniques: Expressing, Western Blot, Isolation, Affinity Purification, Reverse Transcription Polymerase Chain Reaction, Negative Control, Positive Control, Incubation, Nested PCR

    A. Genomic location of ORF 20 and ORF 50 relative to adjacent genes. Genes are indicated by open boxes, arrow head represents direction of transcription. Dotted lines represent introns. The nucleotide positions representing the location of the predicted TATA box and poly A site for each gene are indicated. The position of primers used for PCR and cDNA priming (ORF 20) are indicated by arrows and nucleotide position. Predicted miRNA binding sequences are indicated by vertical bars in the respective UTRs. B. ORF 20 Lane 1, No RT; Lane 2, cDNA primed with 35 pm primer (250 ng); Lane 3, cDNA primed with 10 pm primer (66ng); Lane 4,: No template control; Lane 5, DNA positive control. Lane 6; Marker, Generuler 100bp C. ORF 50 Lane 1, No RT, Lane 2, cDNA primed with Oligo dT; Lane 3, cDNA primed with random primers; Lane 4, No template control; Lane 5, DNA positive control; Lane 6, Marker, Generuler 1kb D. ORF73 Lane 1, No RT, Lane 2, cDNA primed with Oligo dT; Lane 3, cDNA primed with random primers; Lane 4, No template control; Lane 5, DNA positive control; Lane 6, Marker, Generuler 100bp

    Journal: The Journal of general virology

    Article Title: Ovine herpesvirus-2 encoded microRNAs target virus genes involved in virus latency

    doi: 10.1099/vir.0.059303-0

    Figure Lengend Snippet: A. Genomic location of ORF 20 and ORF 50 relative to adjacent genes. Genes are indicated by open boxes, arrow head represents direction of transcription. Dotted lines represent introns. The nucleotide positions representing the location of the predicted TATA box and poly A site for each gene are indicated. The position of primers used for PCR and cDNA priming (ORF 20) are indicated by arrows and nucleotide position. Predicted miRNA binding sequences are indicated by vertical bars in the respective UTRs. B. ORF 20 Lane 1, No RT; Lane 2, cDNA primed with 35 pm primer (250 ng); Lane 3, cDNA primed with 10 pm primer (66ng); Lane 4,: No template control; Lane 5, DNA positive control. Lane 6; Marker, Generuler 100bp C. ORF 50 Lane 1, No RT, Lane 2, cDNA primed with Oligo dT; Lane 3, cDNA primed with random primers; Lane 4, No template control; Lane 5, DNA positive control; Lane 6, Marker, Generuler 1kb D. ORF73 Lane 1, No RT, Lane 2, cDNA primed with Oligo dT; Lane 3, cDNA primed with random primers; Lane 4, No template control; Lane 5, DNA positive control; Lane 6, Marker, Generuler 100bp

    Article Snippet: 1 μg of RNA was digested with RQ1 Dnase (Promega) for 30 min at 37 °C. cDNA was primed using 250ng Oligo dT primer (Promega; equivalent to 0.5 μg/μg RNA) and synthesized using AMV reverse transcriptase for 1 hr at 42 °C.

    Techniques: Polymerase Chain Reaction, Binding Assay, Positive Control, Marker

    : localisation of mRNA. (A) Fluorescent in situ hybridisation assay using specific oligo (dT) probe. As the control, RNase A was incubated with the parasites before probe hybridisation. DAPI: DNA stained with DAPI; mRNA: mRNA detection; granules: labelling of pixels with high intensity fluorescence signals; N: nucleus; K: kinetoplast. Scale bar = 5 µm. (B) Histogram of mRNA granule counts per cell.

    Journal: Memórias do Instituto Oswaldo Cruz

    Article Title: Trypanosoma cruzi transcriptome during axenic epimastigote growth curve

    doi: 10.1590/0074-02760170404

    Figure Lengend Snippet: : localisation of mRNA. (A) Fluorescent in situ hybridisation assay using specific oligo (dT) probe. As the control, RNase A was incubated with the parasites before probe hybridisation. DAPI: DNA stained with DAPI; mRNA: mRNA detection; granules: labelling of pixels with high intensity fluorescence signals; N: nucleus; K: kinetoplast. Scale bar = 5 µm. (B) Histogram of mRNA granule counts per cell.

    Article Snippet: Subsequently, RNA integrity and quality was analysed using the Agilent 2100 bioanalyzer and the RNA 6000 Pico Kit (Agilent). mRNA was selected by oligo-dT chromatography using the PolyATtract® mRNA Isolation Kit (Promega).

    Techniques: In Situ, Hybridization, Incubation, Staining, Fluorescence