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  • 99
    Thermo Fisher mmlv reverse transcriptase
    Determination of the transcription initiation site in the Manduca sexta PAP-3 gene. A primer, complementary to nucleotides 28–56 of the PAP-3 coding region near the 3′ end of exon 1, was terminally labelled with γ- 32 P-dATP and annealed to the total <t>RNA</t> from fat bodies from bacteria-injected larvae (15 µg). After annealing to RNA, the primer was extended with <t>MMLV</t> reverse transcriptase. The set of sequencing reactions (ACGT) on the left of the primer extension lane for use as a sizing ladder was from dideoxynucleotide sequencing of single-stranded M13 mp18 DNA using −40 primer. The arrow indicates the 125 bp extension product from fat body RNA isolated from the induced larvae.
    Mmlv Reverse Transcriptase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6613 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Millipore mmlv reverse transcriptase
    Determination of the transcription initiation site in the Manduca sexta PAP-3 gene. A primer, complementary to nucleotides 28–56 of the PAP-3 coding region near the 3′ end of exon 1, was terminally labelled with γ- 32 P-dATP and annealed to the total <t>RNA</t> from fat bodies from bacteria-injected larvae (15 µg). After annealing to RNA, the primer was extended with <t>MMLV</t> reverse transcriptase. The set of sequencing reactions (ACGT) on the left of the primer extension lane for use as a sizing ladder was from dideoxynucleotide sequencing of single-stranded M13 mp18 DNA using −40 primer. The arrow indicates the 125 bp extension product from fat body RNA isolated from the induced larvae.
    Mmlv Reverse Transcriptase, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 127 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 98 stars, based on 127 article reviews
    Price from $9.99 to $1999.99
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    99
    Promega mmlv reverse transcriptase
    Control Experiments with Recombinant A3A. ( A ) Purification of rA3A from E. coli by Ni affinity: SDS 4–12% polyacrylamide gel electrophoresis and Coomassie blue staining were used to monitor purification of the His-tagged recombinant A3A (rA3A) protein. Input bacterial lysate was loaded onto the Ni-Sepharose column (Lysate). Flow through lysate and wash fractions were collected (Unbound and Wash, respectively). Recombinant A3A was eluted from the column with lysis buffer containing 0.5M imidazole and consecutive elution fractions were collected (F1-4). Most of the His-tagged rA3A was eluted in F2 (arrow). Approximate molecular sizes (kD) are indicated at the left of the gel. ( B ) Purification of rA3A by gel filtration: recombinant A3A purified by Ni-Affinity was further purified by gel filtration by fast protein liquid chromatography (FPLC) on a Superdex 200 column. Arrows indicate the approximate molecular weights (kD) of the proteins. The x-axis indicates the elution volumes and fraction numbers. The y-axis indicates the UV absorbance at 280 nm. ( C ) Recombinant A3A does not deaminate double-stranded DNA: single-strand (lanes 1 and 7) or double-stranded DNA (lanes 2–4 and 8–10) substrates were incubated without (−) or with (+) rA3A (250 ng), were treated with UDG, and the products were resolved by gel electrophoresis on 15% polyacrylamide TBE-Urea Novex gels (Invitrogen). The relative ratios of the target (Oligo) and complementary (asOligo) oligonucleotides are indicated at the top of the figure. A non-specific oligonucleotide (ns) was also included as a control (lanes 6 and 12). As an additional control to rule out potential competition of free asOligo for UDG activity, asOligo was added after rA3A incubation with ssDNA Oligo (lanes 5 and 11). ( D ) Recombinant A3A does not inhibit <t>MMLV-RT</t> activity: from left to right: MMLV RT reactions using purified <t>RNA</t> isolated from pDK101 (WT) or pDK135 (RT-) RNPs, untransfected HeLa cell RNPs (HeLa), and a no RNA sample. Recombinant WT rA3A (100 ng and 300 ng), deaminase-deficient rA3A_C106S (300 ng), and ‘heat-killed’ rA3A (300 ng) were included in MMLV RT reactions. Size standards (bp) are indicated at the left of the gel image. DOI: http://dx.doi.org/10.7554/eLife.02008.006
    Mmlv Reverse Transcriptase, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 6984 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mmlv reverse transcriptase/product/Promega
    Average 99 stars, based on 6984 article reviews
    Price from $9.99 to $1999.99
    mmlv reverse transcriptase - by Bioz Stars, 2020-07
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    99
    TaKaRa m mlv reverse transcriptase
    (A) Laser-scanning confocal microscope was used to detect the distribution of ZIP4 in HepG2, BEL7402, and HL-7702 cells (magnification: ×1260). Cells were incubated in a cell culture dish, fixed with paraformaldehyde for 30 min, and permeabilized with 0.1% Triton X-100 for 5 min at room temperature and incubated with primary polyclonal rabbit anti-human ZIP4 antibody. Cells were then washed and incubated with Alexa Fluor 488 donkey anti-rabbit IgG. The green signal represents staining for ZIP4 protein. ZIP4 located in cell membranes of HL-7702 cells, but not in nuclei. ZIP4 accumulated in the nuclei in HepG2 and BEL7402 cells. (B) The mRNA levels of ZIP4 in seven liver cell lines. We used the <t>TRIzol</t> reagent (Invitrogen) to extract total RNA and performed cDNA synthesis using <t>M-MLV</t> reverse transcriptase (TaKaRa, Dalian, China). ß-actin was the endogenous control. All the samples were normalized to human ß-actin according to the 2 -ΔΔCT method. ZIP4 mRNA levels in immortalized liver cells (HL-7702) was the negative control. The x- axis represents multiples of mRNA levels in HL-7702 cells. (C) ZIP4 protein levels in eight liver cell lines. (D) The mRNA levels of ZIP4 in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. β-actin was used as an internal control. (E) ZIP4 protein levels in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. (F) ZIP4 mRNA levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and negative control (NEG). β-actin was used as an internal control. (G) ZIP4 protein levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and a negative control (NEG). Data are from one of three repeated independent experiments. * P
    M Mlv Reverse Transcriptase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 4323 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Determination of the transcription initiation site in the Manduca sexta PAP-3 gene. A primer, complementary to nucleotides 28–56 of the PAP-3 coding region near the 3′ end of exon 1, was terminally labelled with γ- 32 P-dATP and annealed to the total RNA from fat bodies from bacteria-injected larvae (15 µg). After annealing to RNA, the primer was extended with MMLV reverse transcriptase. The set of sequencing reactions (ACGT) on the left of the primer extension lane for use as a sizing ladder was from dideoxynucleotide sequencing of single-stranded M13 mp18 DNA using −40 primer. The arrow indicates the 125 bp extension product from fat body RNA isolated from the induced larvae.

    Journal: Insect molecular biology

    Article Title: Gene structure and expression profile of Manduca sexta prophenoloxidase-activating proteinase-3 (PAP-3), an immune protein containing two clip domains

    doi: 10.1111/j.1365-2583.2005.00574.x

    Figure Lengend Snippet: Determination of the transcription initiation site in the Manduca sexta PAP-3 gene. A primer, complementary to nucleotides 28–56 of the PAP-3 coding region near the 3′ end of exon 1, was terminally labelled with γ- 32 P-dATP and annealed to the total RNA from fat bodies from bacteria-injected larvae (15 µg). After annealing to RNA, the primer was extended with MMLV reverse transcriptase. The set of sequencing reactions (ACGT) on the left of the primer extension lane for use as a sizing ladder was from dideoxynucleotide sequencing of single-stranded M13 mp18 DNA using −40 primer. The arrow indicates the 125 bp extension product from fat body RNA isolated from the induced larvae.

    Article Snippet: First-strand cDNA synthesis was performed using 2–4 µg RNA, 10 pmol oligo(dT)17 , and 200 U MMLV reverse transcriptase (Invitrogen Life Technologies) at 37 °C for 1 h. M. sexta ribosomal protein S3 transcripts were used as an internal standard to control the template amount in a preliminary PCR experiment.

    Techniques: Injection, Sequencing, Isolation

    Control Experiments with Recombinant A3A. ( A ) Purification of rA3A from E. coli by Ni affinity: SDS 4–12% polyacrylamide gel electrophoresis and Coomassie blue staining were used to monitor purification of the His-tagged recombinant A3A (rA3A) protein. Input bacterial lysate was loaded onto the Ni-Sepharose column (Lysate). Flow through lysate and wash fractions were collected (Unbound and Wash, respectively). Recombinant A3A was eluted from the column with lysis buffer containing 0.5M imidazole and consecutive elution fractions were collected (F1-4). Most of the His-tagged rA3A was eluted in F2 (arrow). Approximate molecular sizes (kD) are indicated at the left of the gel. ( B ) Purification of rA3A by gel filtration: recombinant A3A purified by Ni-Affinity was further purified by gel filtration by fast protein liquid chromatography (FPLC) on a Superdex 200 column. Arrows indicate the approximate molecular weights (kD) of the proteins. The x-axis indicates the elution volumes and fraction numbers. The y-axis indicates the UV absorbance at 280 nm. ( C ) Recombinant A3A does not deaminate double-stranded DNA: single-strand (lanes 1 and 7) or double-stranded DNA (lanes 2–4 and 8–10) substrates were incubated without (−) or with (+) rA3A (250 ng), were treated with UDG, and the products were resolved by gel electrophoresis on 15% polyacrylamide TBE-Urea Novex gels (Invitrogen). The relative ratios of the target (Oligo) and complementary (asOligo) oligonucleotides are indicated at the top of the figure. A non-specific oligonucleotide (ns) was also included as a control (lanes 6 and 12). As an additional control to rule out potential competition of free asOligo for UDG activity, asOligo was added after rA3A incubation with ssDNA Oligo (lanes 5 and 11). ( D ) Recombinant A3A does not inhibit MMLV-RT activity: from left to right: MMLV RT reactions using purified RNA isolated from pDK101 (WT) or pDK135 (RT-) RNPs, untransfected HeLa cell RNPs (HeLa), and a no RNA sample. Recombinant WT rA3A (100 ng and 300 ng), deaminase-deficient rA3A_C106S (300 ng), and ‘heat-killed’ rA3A (300 ng) were included in MMLV RT reactions. Size standards (bp) are indicated at the left of the gel image. DOI: http://dx.doi.org/10.7554/eLife.02008.006

    Journal: eLife

    Article Title: APOBEC3A deaminates transiently exposed single-strand DNA during LINE-1 retrotransposition

    doi: 10.7554/eLife.02008

    Figure Lengend Snippet: Control Experiments with Recombinant A3A. ( A ) Purification of rA3A from E. coli by Ni affinity: SDS 4–12% polyacrylamide gel electrophoresis and Coomassie blue staining were used to monitor purification of the His-tagged recombinant A3A (rA3A) protein. Input bacterial lysate was loaded onto the Ni-Sepharose column (Lysate). Flow through lysate and wash fractions were collected (Unbound and Wash, respectively). Recombinant A3A was eluted from the column with lysis buffer containing 0.5M imidazole and consecutive elution fractions were collected (F1-4). Most of the His-tagged rA3A was eluted in F2 (arrow). Approximate molecular sizes (kD) are indicated at the left of the gel. ( B ) Purification of rA3A by gel filtration: recombinant A3A purified by Ni-Affinity was further purified by gel filtration by fast protein liquid chromatography (FPLC) on a Superdex 200 column. Arrows indicate the approximate molecular weights (kD) of the proteins. The x-axis indicates the elution volumes and fraction numbers. The y-axis indicates the UV absorbance at 280 nm. ( C ) Recombinant A3A does not deaminate double-stranded DNA: single-strand (lanes 1 and 7) or double-stranded DNA (lanes 2–4 and 8–10) substrates were incubated without (−) or with (+) rA3A (250 ng), were treated with UDG, and the products were resolved by gel electrophoresis on 15% polyacrylamide TBE-Urea Novex gels (Invitrogen). The relative ratios of the target (Oligo) and complementary (asOligo) oligonucleotides are indicated at the top of the figure. A non-specific oligonucleotide (ns) was also included as a control (lanes 6 and 12). As an additional control to rule out potential competition of free asOligo for UDG activity, asOligo was added after rA3A incubation with ssDNA Oligo (lanes 5 and 11). ( D ) Recombinant A3A does not inhibit MMLV-RT activity: from left to right: MMLV RT reactions using purified RNA isolated from pDK101 (WT) or pDK135 (RT-) RNPs, untransfected HeLa cell RNPs (HeLa), and a no RNA sample. Recombinant WT rA3A (100 ng and 300 ng), deaminase-deficient rA3A_C106S (300 ng), and ‘heat-killed’ rA3A (300 ng) were included in MMLV RT reactions. Size standards (bp) are indicated at the left of the gel image. DOI: http://dx.doi.org/10.7554/eLife.02008.006

    Article Snippet: RT-PCR was carried out on 0.5 μl of purified RNA, using MMLV reverse transcriptase (Promega, Madison, WI) and 0.8 μM LEAP adapter (5np1) at 42°C for 30 min.

    Techniques: Recombinant, Purification, Polyacrylamide Gel Electrophoresis, Staining, Flow Cytometry, Lysis, Filtration, Fast Protein Liquid Chromatography, Incubation, Nucleic Acid Electrophoresis, Activity Assay, Isolation

    (A) Laser-scanning confocal microscope was used to detect the distribution of ZIP4 in HepG2, BEL7402, and HL-7702 cells (magnification: ×1260). Cells were incubated in a cell culture dish, fixed with paraformaldehyde for 30 min, and permeabilized with 0.1% Triton X-100 for 5 min at room temperature and incubated with primary polyclonal rabbit anti-human ZIP4 antibody. Cells were then washed and incubated with Alexa Fluor 488 donkey anti-rabbit IgG. The green signal represents staining for ZIP4 protein. ZIP4 located in cell membranes of HL-7702 cells, but not in nuclei. ZIP4 accumulated in the nuclei in HepG2 and BEL7402 cells. (B) The mRNA levels of ZIP4 in seven liver cell lines. We used the TRIzol reagent (Invitrogen) to extract total RNA and performed cDNA synthesis using M-MLV reverse transcriptase (TaKaRa, Dalian, China). ß-actin was the endogenous control. All the samples were normalized to human ß-actin according to the 2 -ΔΔCT method. ZIP4 mRNA levels in immortalized liver cells (HL-7702) was the negative control. The x- axis represents multiples of mRNA levels in HL-7702 cells. (C) ZIP4 protein levels in eight liver cell lines. (D) The mRNA levels of ZIP4 in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. β-actin was used as an internal control. (E) ZIP4 protein levels in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. (F) ZIP4 mRNA levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and negative control (NEG). β-actin was used as an internal control. (G) ZIP4 protein levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and a negative control (NEG). Data are from one of three repeated independent experiments. * P

    Journal: International Journal of Biological Sciences

    Article Title: ZIP4, a Novel Determinant of Tumor Invasion in Hepatocellular Carcinoma, Contributes to Tumor Recurrence after Liver Transplantation

    doi: 10.7150/ijbs.7401

    Figure Lengend Snippet: (A) Laser-scanning confocal microscope was used to detect the distribution of ZIP4 in HepG2, BEL7402, and HL-7702 cells (magnification: ×1260). Cells were incubated in a cell culture dish, fixed with paraformaldehyde for 30 min, and permeabilized with 0.1% Triton X-100 for 5 min at room temperature and incubated with primary polyclonal rabbit anti-human ZIP4 antibody. Cells were then washed and incubated with Alexa Fluor 488 donkey anti-rabbit IgG. The green signal represents staining for ZIP4 protein. ZIP4 located in cell membranes of HL-7702 cells, but not in nuclei. ZIP4 accumulated in the nuclei in HepG2 and BEL7402 cells. (B) The mRNA levels of ZIP4 in seven liver cell lines. We used the TRIzol reagent (Invitrogen) to extract total RNA and performed cDNA synthesis using M-MLV reverse transcriptase (TaKaRa, Dalian, China). ß-actin was the endogenous control. All the samples were normalized to human ß-actin according to the 2 -ΔΔCT method. ZIP4 mRNA levels in immortalized liver cells (HL-7702) was the negative control. The x- axis represents multiples of mRNA levels in HL-7702 cells. (C) ZIP4 protein levels in eight liver cell lines. (D) The mRNA levels of ZIP4 in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. β-actin was used as an internal control. (E) ZIP4 protein levels in BEL7402 and HepG2 cells with ZIP4 silencing (shRNA-ZIP4) and negative control (NC) cells. (F) ZIP4 mRNA levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and negative control (NEG). β-actin was used as an internal control. (G) ZIP4 protein levels in HuH-7 and HepG2 cells with ZIP4 overexpression (Z1191) and a negative control (NEG). Data are from one of three repeated independent experiments. * P

    Article Snippet: Real-time polymerase chain reaction RNA was extracted using the TRIzol reagent (Invitrogen) and used for cDNA synthesis with a M-MLV Reverse Transcriptase (TaKaRa, Dalian, China).

    Techniques: Microscopy, Incubation, Cell Culture, Staining, Negative Control, shRNA, Over Expression