pcr clean up  (Thermo Fisher)


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    Name:
    ExoSAP IT Express PCR Product Cleanup Reagent
    Description:
    In only 5 minutes ExoSAP IT Express PCR Product Cleanup Reagent removes excess primers and unincorporated nucleotides from a PCR reaction This enzymatic cleanup method offers precision increased yield and a faster turnaround time compared to alternative purification methods such as spin columns or beads • Remove unincorporated primers and nucleotides in about 5 minutes • Prepare DNA sequencing samples with one pipetting step • Conserve precious PCR products with up to 100 recovery regardless of amplicon length • Minimize spin column or bead cleanup ExoSAP IT Express PCR Product Cleanup Reagent consists of two hydrolytic enzymes a novel exonuclease I and shrimp alkaline phosphatase SAP in a specially formulated buffer The reaction setup is complete with one pipetting step which is followed by two short incubations The first incubation hydrolyzes excess primer and dephosphorylates nucleotides The second high temperature incubation completely and irreversibly inactivates the enzymes in about one minute Adding ExoSAP IT Express reagent directly to the PCR product eliminates transfer steps to tubes wells or columns conserves PCR amplicons helping reduce the chance of cross contamination no further processing is needed Samples are ready for DNA sequencing or SNP analysis
    Catalog Number:
    75001.1.ea
    Price:
    None
    Applications:
    PCR|PCR & Real-Time PCR|Sequencing
    Category:
    Proteins Enzymes Peptides
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    Structured Review

    Thermo Fisher pcr clean up
    In only 5 minutes ExoSAP IT Express PCR Product Cleanup Reagent removes excess primers and unincorporated nucleotides from a PCR reaction This enzymatic cleanup method offers precision increased yield and a faster turnaround time compared to alternative purification methods such as spin columns or beads • Remove unincorporated primers and nucleotides in about 5 minutes • Prepare DNA sequencing samples with one pipetting step • Conserve precious PCR products with up to 100 recovery regardless of amplicon length • Minimize spin column or bead cleanup ExoSAP IT Express PCR Product Cleanup Reagent consists of two hydrolytic enzymes a novel exonuclease I and shrimp alkaline phosphatase SAP in a specially formulated buffer The reaction setup is complete with one pipetting step which is followed by two short incubations The first incubation hydrolyzes excess primer and dephosphorylates nucleotides The second high temperature incubation completely and irreversibly inactivates the enzymes in about one minute Adding ExoSAP IT Express reagent directly to the PCR product eliminates transfer steps to tubes wells or columns conserves PCR amplicons helping reduce the chance of cross contamination no further processing is needed Samples are ready for DNA sequencing or SNP analysis
    https://www.bioz.com/result/pcr clean up/product/Thermo Fisher
    Average 99 stars, based on 31 article reviews
    Price from $9.99 to $1999.99
    pcr clean up - by Bioz Stars, 2020-08
    99/100 stars

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    Related Articles

    Amplification:

    Article Title: Molecular evidence supports the expansion of visceral leishmaniasis towards non-program districts of Nepal
    Article Snippet: .. Amplicon purification and sequencing The PCR product was purified using ExoSAP-IT Express (Affymetrix, Inc., CA, USA) according to the manufacturer’s manual. .. For this, 5 μl of PCR product and 2 μl of Exosap was mixed and incubated at 37 °C for 4 min followed by 80 °C for 1 min.

    Agarose Gel Electrophoresis:

    Article Title: GATA4 mediates gene repression in the mature mouse small intestine through interactions with Friend of GATA (FOG) cofactors
    Article Snippet: .. The PCR product was separated on an agarose gel by electrophoresis, extracted using the QIAquick Gel Extraction Kit (Qiagen), re-amplified using the same primers, and purified using ExoSap-IT (USB Corporation). .. The purified PCR product was sequenced at the Molecular Genetics Core (Children’s Hospital Boston) using a nested primer.

    Purification:

    Article Title: Disrupted auto-regulation of the spliceosomal gene SNRPB causes cerebro–costo–mandibular syndrome
    Article Snippet: .. A quantity of 1.2 μl of 1/20 dilution of the PCR product was purified in a reaction with 1 μl ExoSAP-IT (Affymetrix) and 3 μl H2 O The product of this reaction was added to a sequencing reaction with 2.2 μl H2 O, 1.875 μl of 5 × sequencing buffer, 0.5 μl primer and 0.25 μl BigDye Terminator v1.1 (Life Technologies). .. Unincorporated nucleotides were removed from the sequencing reaction by passage through a Sephadex column.

    Article Title: Molecular evidence supports the expansion of visceral leishmaniasis towards non-program districts of Nepal
    Article Snippet: .. Amplicon purification and sequencing The PCR product was purified using ExoSAP-IT Express (Affymetrix, Inc., CA, USA) according to the manufacturer’s manual. .. For this, 5 μl of PCR product and 2 μl of Exosap was mixed and incubated at 37 °C for 4 min followed by 80 °C for 1 min.

    Article Title: Accurate detection of KRAS, NRAS and BRAF mutations in metastatic colorectal cancers by bridged nucleic acid-clamp real-time PCR
    Article Snippet: .. BNA-clamp PCR products were purified using ExoSAP-IT Express PCR Cleanup Reagent (Thermo Fisher Scientific) [ , ]. .. Purified products were used as templates and Sanger sequencing was performed using the BNA Real-time PCR Extended RAS Mutation Sequencing Primer (Riken Genesis) and the BigDye® Terminator v3.1 Cycle Sequencing Kit (Thermo Fisher Scientific).

    Article Title: GATA4 mediates gene repression in the mature mouse small intestine through interactions with Friend of GATA (FOG) cofactors
    Article Snippet: .. The PCR product was separated on an agarose gel by electrophoresis, extracted using the QIAquick Gel Extraction Kit (Qiagen), re-amplified using the same primers, and purified using ExoSap-IT (USB Corporation). .. The purified PCR product was sequenced at the Molecular Genetics Core (Children’s Hospital Boston) using a nested primer.

    Electrophoresis:

    Article Title: GATA4 mediates gene repression in the mature mouse small intestine through interactions with Friend of GATA (FOG) cofactors
    Article Snippet: .. The PCR product was separated on an agarose gel by electrophoresis, extracted using the QIAquick Gel Extraction Kit (Qiagen), re-amplified using the same primers, and purified using ExoSap-IT (USB Corporation). .. The purified PCR product was sequenced at the Molecular Genetics Core (Children’s Hospital Boston) using a nested primer.

    Polymerase Chain Reaction:

    Article Title: Accurate detection of de novo and transmitted indels within exome-capture data using micro-assembly
    Article Snippet: .. PCR product was verified on E-Gel® 96 gels (Invitrogen Corp., Carlsbad, CA, USA) and subsequently pooled for ExoSAP-IT® (Affymetrix Inc., Santa Clara, CA, USA) cleanup. .. The cleanup product was further purified using QIAquick PCR Purification Kit (QIAGEN Inc., Valencia, CA, USA) and quantified by Qubit® dsDNA BR Assay Kit (Invitrogen Corp.).

    Article Title: Disrupted auto-regulation of the spliceosomal gene SNRPB causes cerebro–costo–mandibular syndrome
    Article Snippet: .. A quantity of 1.2 μl of 1/20 dilution of the PCR product was purified in a reaction with 1 μl ExoSAP-IT (Affymetrix) and 3 μl H2 O The product of this reaction was added to a sequencing reaction with 2.2 μl H2 O, 1.875 μl of 5 × sequencing buffer, 0.5 μl primer and 0.25 μl BigDye Terminator v1.1 (Life Technologies). .. Unincorporated nucleotides were removed from the sequencing reaction by passage through a Sephadex column.

    Article Title: Extended-Spectrum Beta-Lactamase Gene Sequences in Gram-Negative Saprophytes on Retail Organic and Nonorganic Spinach ▿
    Article Snippet: .. Each PCR product that showed an electrophoretic band of an expected size was cleaned up by using ExoSAP-IT according to the manufacturer's instructions (USB Corporation). ..

    Article Title: Molecular evidence supports the expansion of visceral leishmaniasis towards non-program districts of Nepal
    Article Snippet: .. Amplicon purification and sequencing The PCR product was purified using ExoSAP-IT Express (Affymetrix, Inc., CA, USA) according to the manufacturer’s manual. .. For this, 5 μl of PCR product and 2 μl of Exosap was mixed and incubated at 37 °C for 4 min followed by 80 °C for 1 min.

    Article Title: Accurate detection of KRAS, NRAS and BRAF mutations in metastatic colorectal cancers by bridged nucleic acid-clamp real-time PCR
    Article Snippet: .. BNA-clamp PCR products were purified using ExoSAP-IT Express PCR Cleanup Reagent (Thermo Fisher Scientific) [ , ]. .. Purified products were used as templates and Sanger sequencing was performed using the BNA Real-time PCR Extended RAS Mutation Sequencing Primer (Riken Genesis) and the BigDye® Terminator v3.1 Cycle Sequencing Kit (Thermo Fisher Scientific).

    Article Title: C3orf70 Is Involved in Neural and Neurobehavioral Development
    Article Snippet: .. The PCR product corresponding to the homo-KO of c3orf70a or c3orf70b was subjected to sequence analysis using ExoSAP-IT Express PCR Cleanup Reagents (Thermo Fisher, MA, USA) according to the manufacturer’s protocol. .. F2 fish (c3orf70a+/- :c3orf70b+/ -) with a homozygous genotype (validated to have the frame shift mutation, as shown in ) were used to generate F3 progeny (c3orf70a-/- :c3orf70b-/- ).

    Article Title: New susceptibility locus for obesity and dyslipidaemia on chromosome 3q22.3
    Article Snippet: .. Five microlitres of PCR product was then treated with 2 μl of ExoSAP-IT (USB Corporation, Cleveland, OH, USA) at 37°C for 30 min to allow the hydrolytic removal of excess primers by exonuclease 1 and shrimp alkaline phosphatase. .. The enzymes were inactivated at 80°C for 15 min, and the sequencing reaction was initiated by mixing 2 μl of DNA, 1 μl of 5 μmol primer, 8 μl of DYEnamic ET Dye Terminator (Amersham Biosciences, Amersham, Buckinghamshire, UK) and 9 μl of distilled water.

    Article Title: GATA4 mediates gene repression in the mature mouse small intestine through interactions with Friend of GATA (FOG) cofactors
    Article Snippet: .. The PCR product was separated on an agarose gel by electrophoresis, extracted using the QIAquick Gel Extraction Kit (Qiagen), re-amplified using the same primers, and purified using ExoSap-IT (USB Corporation). .. The purified PCR product was sequenced at the Molecular Genetics Core (Children’s Hospital Boston) using a nested primer.

    Sequencing:

    Article Title: Disrupted auto-regulation of the spliceosomal gene SNRPB causes cerebro–costo–mandibular syndrome
    Article Snippet: .. A quantity of 1.2 μl of 1/20 dilution of the PCR product was purified in a reaction with 1 μl ExoSAP-IT (Affymetrix) and 3 μl H2 O The product of this reaction was added to a sequencing reaction with 2.2 μl H2 O, 1.875 μl of 5 × sequencing buffer, 0.5 μl primer and 0.25 μl BigDye Terminator v1.1 (Life Technologies). .. Unincorporated nucleotides were removed from the sequencing reaction by passage through a Sephadex column.

    Article Title: Molecular evidence supports the expansion of visceral leishmaniasis towards non-program districts of Nepal
    Article Snippet: .. Amplicon purification and sequencing The PCR product was purified using ExoSAP-IT Express (Affymetrix, Inc., CA, USA) according to the manufacturer’s manual. .. For this, 5 μl of PCR product and 2 μl of Exosap was mixed and incubated at 37 °C for 4 min followed by 80 °C for 1 min.

    Article Title: C3orf70 Is Involved in Neural and Neurobehavioral Development
    Article Snippet: .. The PCR product corresponding to the homo-KO of c3orf70a or c3orf70b was subjected to sequence analysis using ExoSAP-IT Express PCR Cleanup Reagents (Thermo Fisher, MA, USA) according to the manufacturer’s protocol. .. F2 fish (c3orf70a+/- :c3orf70b+/ -) with a homozygous genotype (validated to have the frame shift mutation, as shown in ) were used to generate F3 progeny (c3orf70a-/- :c3orf70b-/- ).

    Gel Extraction:

    Article Title: GATA4 mediates gene repression in the mature mouse small intestine through interactions with Friend of GATA (FOG) cofactors
    Article Snippet: .. The PCR product was separated on an agarose gel by electrophoresis, extracted using the QIAquick Gel Extraction Kit (Qiagen), re-amplified using the same primers, and purified using ExoSap-IT (USB Corporation). .. The purified PCR product was sequenced at the Molecular Genetics Core (Children’s Hospital Boston) using a nested primer.

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  • 92
    Thermo Fisher pcr clean up kit
    Essential role of PRAMEF12 in spermatogonial differentiation. a Immunofluorescence of whole-mount testes from P2, P4, and P7 Pramef12 Null and Pramef12 Het mice after staining with antibodies to DDX4 (left) or KIT (middle) and merged (right). KIT-positive spermatogonia were rarely detected at P2 in either Pramef12 Null or Pramef12 Het tubules but were present at P4 and increased at P7 in Pramef12 Het tubules. Few KIT-positive spermatogonia (arrow) were observed in P7 Pramef12 Null tubules. Scale bar, 50 μm. b Immunohistochemistry of sections from P7 Pramef12 Null (top panels) and Pramef12 Het (bottom panels) testes after staining with antibodies to DDX4 (left) or KIT (middle) and merged with Hoechst 33342 to stain DNA (right). Arrowheads, KIT-positive cells in Pramef12 Null testes. Scale bar, 50 μm. c Quantitative real-time <t>RT-PCR</t> of Kit <t>mRNA</t> abundance in P7 Pramef12 Null and Pramef12 Het testes using β-actin as an internal load control and setting the abundance in Pramef12 Het testis to 1. Mean ± s.d, n = 3 biologically independent samples per condition. * P = 1.43E-05 by two-tailed Student’s t test. d Immunoblot of KIT protein in P7 Pramef12 Null and Pramef12 Het testes using α-tubulin as a load control. e Same as a , but with antibodies to SOHLH1, SOHLH2, and PLZF. The number of spermatogonia positive for SOHLH1 and SOHLH2 was significantly reduced in P7 Pramef12 Null tubules (top panels). Representative of n = 3 a , b , d , e independent biological replicates with similar results per condition
    Pcr Clean Up Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 35 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr clean up kit/product/Thermo Fisher
    Average 92 stars, based on 35 article reviews
    Price from $9.99 to $1999.99
    pcr clean up kit - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    84
    Thermo Fisher exosap pcr clean up reagent
    Essential role of PRAMEF12 in spermatogonial differentiation. a Immunofluorescence of whole-mount testes from P2, P4, and P7 Pramef12 Null and Pramef12 Het mice after staining with antibodies to DDX4 (left) or KIT (middle) and merged (right). KIT-positive spermatogonia were rarely detected at P2 in either Pramef12 Null or Pramef12 Het tubules but were present at P4 and increased at P7 in Pramef12 Het tubules. Few KIT-positive spermatogonia (arrow) were observed in P7 Pramef12 Null tubules. Scale bar, 50 μm. b Immunohistochemistry of sections from P7 Pramef12 Null (top panels) and Pramef12 Het (bottom panels) testes after staining with antibodies to DDX4 (left) or KIT (middle) and merged with Hoechst 33342 to stain DNA (right). Arrowheads, KIT-positive cells in Pramef12 Null testes. Scale bar, 50 μm. c Quantitative real-time <t>RT-PCR</t> of Kit <t>mRNA</t> abundance in P7 Pramef12 Null and Pramef12 Het testes using β-actin as an internal load control and setting the abundance in Pramef12 Het testis to 1. Mean ± s.d, n = 3 biologically independent samples per condition. * P = 1.43E-05 by two-tailed Student’s t test. d Immunoblot of KIT protein in P7 Pramef12 Null and Pramef12 Het testes using α-tubulin as a load control. e Same as a , but with antibodies to SOHLH1, SOHLH2, and PLZF. The number of spermatogonia positive for SOHLH1 and SOHLH2 was significantly reduced in P7 Pramef12 Null tubules (top panels). Representative of n = 3 a , b , d , e independent biological replicates with similar results per condition
    Exosap Pcr Clean Up Reagent, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/exosap pcr clean up reagent/product/Thermo Fisher
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    exosap pcr clean up reagent - by Bioz Stars, 2020-08
    84/100 stars
      Buy from Supplier

    90
    Thermo Fisher pcr clean up kit machery nagel
    Essential role of PRAMEF12 in spermatogonial differentiation. a Immunofluorescence of whole-mount testes from P2, P4, and P7 Pramef12 Null and Pramef12 Het mice after staining with antibodies to DDX4 (left) or KIT (middle) and merged (right). KIT-positive spermatogonia were rarely detected at P2 in either Pramef12 Null or Pramef12 Het tubules but were present at P4 and increased at P7 in Pramef12 Het tubules. Few KIT-positive spermatogonia (arrow) were observed in P7 Pramef12 Null tubules. Scale bar, 50 μm. b Immunohistochemistry of sections from P7 Pramef12 Null (top panels) and Pramef12 Het (bottom panels) testes after staining with antibodies to DDX4 (left) or KIT (middle) and merged with Hoechst 33342 to stain DNA (right). Arrowheads, KIT-positive cells in Pramef12 Null testes. Scale bar, 50 μm. c Quantitative real-time <t>RT-PCR</t> of Kit <t>mRNA</t> abundance in P7 Pramef12 Null and Pramef12 Het testes using β-actin as an internal load control and setting the abundance in Pramef12 Het testis to 1. Mean ± s.d, n = 3 biologically independent samples per condition. * P = 1.43E-05 by two-tailed Student’s t test. d Immunoblot of KIT protein in P7 Pramef12 Null and Pramef12 Het testes using α-tubulin as a load control. e Same as a , but with antibodies to SOHLH1, SOHLH2, and PLZF. The number of spermatogonia positive for SOHLH1 and SOHLH2 was significantly reduced in P7 Pramef12 Null tubules (top panels). Representative of n = 3 a , b , d , e independent biological replicates with similar results per condition
    Pcr Clean Up Kit Machery Nagel, 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/result/pcr clean up kit machery nagel/product/Thermo Fisher
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pcr clean up kit machery nagel - by Bioz Stars, 2020-08
    90/100 stars
      Buy from Supplier

    Image Search Results


    Essential role of PRAMEF12 in spermatogonial differentiation. a Immunofluorescence of whole-mount testes from P2, P4, and P7 Pramef12 Null and Pramef12 Het mice after staining with antibodies to DDX4 (left) or KIT (middle) and merged (right). KIT-positive spermatogonia were rarely detected at P2 in either Pramef12 Null or Pramef12 Het tubules but were present at P4 and increased at P7 in Pramef12 Het tubules. Few KIT-positive spermatogonia (arrow) were observed in P7 Pramef12 Null tubules. Scale bar, 50 μm. b Immunohistochemistry of sections from P7 Pramef12 Null (top panels) and Pramef12 Het (bottom panels) testes after staining with antibodies to DDX4 (left) or KIT (middle) and merged with Hoechst 33342 to stain DNA (right). Arrowheads, KIT-positive cells in Pramef12 Null testes. Scale bar, 50 μm. c Quantitative real-time RT-PCR of Kit mRNA abundance in P7 Pramef12 Null and Pramef12 Het testes using β-actin as an internal load control and setting the abundance in Pramef12 Het testis to 1. Mean ± s.d, n = 3 biologically independent samples per condition. * P = 1.43E-05 by two-tailed Student’s t test. d Immunoblot of KIT protein in P7 Pramef12 Null and Pramef12 Het testes using α-tubulin as a load control. e Same as a , but with antibodies to SOHLH1, SOHLH2, and PLZF. The number of spermatogonia positive for SOHLH1 and SOHLH2 was significantly reduced in P7 Pramef12 Null tubules (top panels). Representative of n = 3 a , b , d , e independent biological replicates with similar results per condition

    Journal: Nature Communications

    Article Title: Sertoli cell-only phenotype and scRNA-seq define PRAMEF12 as a factor essential for spermatogenesis in mice

    doi: 10.1038/s41467-019-13193-3

    Figure Lengend Snippet: Essential role of PRAMEF12 in spermatogonial differentiation. a Immunofluorescence of whole-mount testes from P2, P4, and P7 Pramef12 Null and Pramef12 Het mice after staining with antibodies to DDX4 (left) or KIT (middle) and merged (right). KIT-positive spermatogonia were rarely detected at P2 in either Pramef12 Null or Pramef12 Het tubules but were present at P4 and increased at P7 in Pramef12 Het tubules. Few KIT-positive spermatogonia (arrow) were observed in P7 Pramef12 Null tubules. Scale bar, 50 μm. b Immunohistochemistry of sections from P7 Pramef12 Null (top panels) and Pramef12 Het (bottom panels) testes after staining with antibodies to DDX4 (left) or KIT (middle) and merged with Hoechst 33342 to stain DNA (right). Arrowheads, KIT-positive cells in Pramef12 Null testes. Scale bar, 50 μm. c Quantitative real-time RT-PCR of Kit mRNA abundance in P7 Pramef12 Null and Pramef12 Het testes using β-actin as an internal load control and setting the abundance in Pramef12 Het testis to 1. Mean ± s.d, n = 3 biologically independent samples per condition. * P = 1.43E-05 by two-tailed Student’s t test. d Immunoblot of KIT protein in P7 Pramef12 Null and Pramef12 Het testes using α-tubulin as a load control. e Same as a , but with antibodies to SOHLH1, SOHLH2, and PLZF. The number of spermatogonia positive for SOHLH1 and SOHLH2 was significantly reduced in P7 Pramef12 Null tubules (top panels). Representative of n = 3 a , b , d , e independent biological replicates with similar results per condition

    Article Snippet: Cas9 mRNA (Addgene #42251) was generated by linearization with PmeI, purified with PCR clean-up kit, and transcribed with mMESSAGE mMACHINE T7 Kit (Thermo Fisher Scientific).

    Techniques: Immunofluorescence, Mouse Assay, Staining, Immunohistochemistry, Quantitative RT-PCR, Two Tailed Test