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Meridian Life Science dntp
Dntp, supplied by Meridian Life Science, used in various techniques. Bioz Stars score: 99/100, based on 278 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 99 stars, based on 278 article reviews
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dntp - by Bioz Stars, 2020-04
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Amplification:

Article Title: Predominant Role of Host Genetics in Controlling the Composition of Gut Microbiota
Article Snippet: Amplification of the 16S rRNA genes was carried out with the universal primer set–27F ( 5′-AGAGTTTGATCMTGGCTCAG-3′ ; positions 8 to 27 in the Escherichia coli 16S rRNA gene) and 1492R ( 5′-ACGGCTACCTTGTTACGACTT-3′ ; positions 1510 to 1492 in the E. coli 16S rRNA gene) . .. The PCR reaction mixture contained 10 mM Tris-HCl, 2 mM MgCl2 , 50 mM KCl, 200 µM of each dNTP, 10.0 pmol of each primer, 1U of Bio-X-Act Short DNA polymerase (Bioline, UK), and 100 ng of template DNA in a final volume of 50 µl.

Article Title: The Colletotrichum acutatum species complex
Article Snippet: The 5.8S nuclear ribosomal gene with the two flanking internal transcribed spacers (ITS), a 200-bp intron of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and partial sequences of the chitin synthase 1 (CHS-1), histone3 (HIS3), actin (ACT) and beta-tubulin (TUB2) genes were amplified and sequenced using the primer pairs ITS-1F ( ) + ITS-4 ( ) or V9G ( ) + ITS-4, GDF1 + GDR1 , CHS-354R + CHS-79F , CYLH3F + CYLH3R , ACT-512F + ACT-783R ( ) and BT2Fd + BT4R ( ) or T1 ( ) + Bt-2b , respectively. .. The GAPDH, CHS-1, HIS3, ACT and TUB2 PCR mixture contained 1 μL 20x diluted genomic DNA, 0.2 μM of each primer, 1x PCR buffer (Bioline, Luckenwalde, Germany), 2 mM MgCl2 , 20 μM of each dNTP, 0.7 μL DMSO and 0.25 U Taq DNA polymerase (Bioline).

Article Title: Assortative mating among Lake Malawi cichlid fish populations is not simply predictable from male nuptial colour
Article Snippet: DNA was extracted from tissue samples using the HOTSHOT method [ ] and amplified using a FAM/HEX/TET-labelled forward primer. .. PCR conditions consisted of 30 cycles of 30 sec at 95°, 60 sec at 55°, 60 sec at 72° in a 20 μl reaction volume containing 0.25 mM of each dNTP, 2 μl of 10× reaction buffer (Bioline, London, UK) 1.5 mM MgCl2 , 0.5 U of taq DNA polymerase (Bioline, London, UK), 25 ρmol of each primer and approximately 1 μg of template genomic DNA in a TGradient thermocycler (Biometra, Göttingen, Germany).

Article Title: Early infant diagnosis of HIV-1 infection in Luanda, Angola, using a new DNA PCR assay and dried blood spots
Article Snippet: Paragraph title: PCR amplification of proviral HIV-1 DNA ... The 25 μL reaction volume contained 1X NH4 buffer, 3 mmol/L MgCl2, 0.5 μmol/L of each dNTP, 0.3 μmol/L forward and reverse primers , 1U of Taq DNA polymerase (Bioline® Reagents Ltd, London, UK) plus 2.5 μL of DNA solution from the clinical or control samples (first-round PCR).

Article Title: Optimizing Production in the New Generation of Apricot Cultivars: Self-incompatibility, S-RNase Allele Identification, and Incompatibility Group Assignment
Article Snippet: S-RNase allele identification by PCR analysis Amplification reactions for the first intron region of the S-RNase gene were carried out with the combination of the fluorescently labeled forward primer SRc-F (5′-CTCGCTTTCCTTGTTCTTGC-3′) with the reverse primer SRc-R (5′-GGCCATTGTTGCACAAATTG-3′; Romero et al., ; Vilanova et al., ). .. PCR amplifications were carried out in 15 μl reaction volumes, containing 10x NH4 Reaction Buffer, 25 mM Cl2 Mg, 2.5 mM of each dNTP, 10 μM of each primer, 100 ng of genomic DNA and 0.5 U of BioTaq™ DNA polymerase (Bioline, London, UK).

Article Title: Multiple Ethnic Origins of Mitochondrial DNA Lineages for the Population of Mauritius
Article Snippet: The complete hypervariable region (HVR) was amplified using the primers L15676 and H00945, previously described by Maca-Meyer et al. . .. The PCR was carried out in 50-μl volumes, containing 1X Tris–HCl buffer, 200 μM of each dNTP, 2.5 mM MgCl2, 50 pmoles of each primer and 3 U of Taq polymerase (Bioline).

Article Title: Modification of a Rodent Hindlimb Model of Secondary Lymphedema: Surgical Radicality versus Radiotherapeutic Ablation
Article Snippet: The denatured RNA was mixed with 4 μ L of 5x reaction buffer, 1 mM dNTP, 20 U of RNase inhibitor, and reverse transcriptase BioScript (Bioline) to make a final volume of 20 μ LcDNA. .. For PCR amplification, cDNA equivalent to 500 ng of starting RNA was mixed with 20 pmol of forward and reverse primers with 10x PCR buffer (MangoMix, Bioline).

Article Title: The prognostic efficacy of cell-free DNA hypermethylation in colorectal cancer
Article Snippet: We added 10 μl of the first round PCR product to 710 μl preincubated reaction mix (37 °C for five minutes and 95 °C for 10 minutes) containing PCR buffer, 250 μM dNTP, 10 μM MgCl2 , 8 U Taq polymerase (Bioline® , [Taunton, MA, USA]), and 0.8 Uracil-DNA Glycosylase (Invitrogen® [Waltham, MA, USA]). .. All primer and probe sequences along with amplicon sizes are available in - .

Article Title: The Use of Fluorescent Fragment Length Analysis (PCR-FFL) in the Direct Diagnosis and Identification of Cutaneous Leishmania Species
Article Snippet: .. Amplification reactions were performed in volumes of 50 μL containing 5 μL of 10X buffer (BIOTAQ DNA Polymerase, Bioline, London, UK), 1.5 mM MgCl2 , 0.2 mM dNTP, 0.2 μM of each primer and 1.5 units of Taq polymerase (BIOTAQ DNA Polymerase, Bioline, London, UK). .. We added 3 μL of isolated DNA to the mixture and incubated it in the thermal cycler (MJ Research PTC-200 DNA Engine, Alameda, CA) under the following conditions: for the ITS-1 PCR, we started with a denaturing step at 95°C for 2 min, followed by 35 cycles of denaturing for 20 sec at 95°C, annealing for 30 sec at 53°C, and extension for 1 min at 72°C, followed by a final extension at 72°C for 1 h. For the 7SL PCR, the denaturing step lasted for 5 min at 95°C, followed by 35 cycles of denaturing for 20 sec at 95°C, annealing for 30 sec at 65°C, and extension for 1 min at 72°C.

Article Title: Association of factor H autoantibodies with deletions of CFHR1, CFHR3, CFHR4, and with mutations in CFH, CFI, CD46, and C3 in patients with atypical hemolytic uremic syndrome
Article Snippet: CFHR1 and CFHR3 copy number was measured by the use of multiplex ligation-dependent probe amplification with a kit from MRC Holland (SALSA MLPA kit P236-A1 ARMD). .. PCRs were carried out in 25-μL volumes with the use of 150 ng of DNA and contained 0.5mM each dNTP, 6.7mM MgCl2 , 12.5pM of each primer, 1 U of a hot-start Taq polymerase (Immolase, Bioline) in a buffer of 16mM (NH4 )2 SO4 , 67mM Tris-HCI, and 0.01% Tween-20.

Mass Spectrometry:

Article Title: Assortative mating among Lake Malawi cichlid fish populations is not simply predictable from male nuptial colour
Article Snippet: 5–10 days after spawning and their offspring removed by gently opening the female's mouth and allowing the fry to drop into a tray of water containing an anaesthetic overdose (MS-222). .. PCR conditions consisted of 30 cycles of 30 sec at 95°, 60 sec at 55°, 60 sec at 72° in a 20 μl reaction volume containing 0.25 mM of each dNTP, 2 μl of 10× reaction buffer (Bioline, London, UK) 1.5 mM MgCl2 , 0.5 U of taq DNA polymerase (Bioline, London, UK), 25 ρmol of each primer and approximately 1 μg of template genomic DNA in a TGradient thermocycler (Biometra, Göttingen, Germany).

Synthesized:

Article Title: Somatostatin and opioid receptors do not regulate proliferation or apoptosis of the human multiple myeloma U266 cells
Article Snippet: RT-PCR Total RNAs were extracted using the RNAgents® Total RNA Isolation System (Promega) according to Chomczynski and Sacchi [ ]. cDNAs were synthesized from 2 μg of RNA in a buffer supplied with the reverse transcriptase (RT) (Promega) containing 900 μM dNTP (Amersham), 20 units RNAsine (Promega), 500 ng random primers (Promega) and 200 units of Moloney murine leukaemia virus RT in a final volume of 20 μL. .. PCRs were performed using 2 μL of cDNAs in the PCR buffer supplied with the Taq polymerase supplemented with 1.5 mM MgCl2 , 0.2 mM of dNTP, 2.5 units of Taq polymerase (Bioline), and 0.5 μM of each sense and antisense primer.

Real-time Polymerase Chain Reaction:

Article Title: The prognostic efficacy of cell-free DNA hypermethylation in colorectal cancer
Article Snippet: We added 10 μl of the first round PCR product to 710 μl preincubated reaction mix (37 °C for five minutes and 95 °C for 10 minutes) containing PCR buffer, 250 μM dNTP, 10 μM MgCl2 , 8 U Taq polymerase (Bioline® , [Taunton, MA, USA]), and 0.8 Uracil-DNA Glycosylase (Invitrogen® [Waltham, MA, USA]). .. Real time PCR was conducted for 45 rounds (94 °C for 15 seconds, 55 °C for 30 seconds, and 72 °C for 30 seconds).

Incubation:

Article Title: Modification of a Rodent Hindlimb Model of Secondary Lymphedema: Surgical Radicality versus Radiotherapeutic Ablation
Article Snippet: An oligonucleotide deoxythymidine primer was added to 2 μ g of the extracted total RNA and incubated at 65°C for 10 min. .. The denatured RNA was mixed with 4 μ L of 5x reaction buffer, 1 mM dNTP, 20 U of RNase inhibitor, and reverse transcriptase BioScript (Bioline) to make a final volume of 20 μ LcDNA.

Article Title: The prognostic efficacy of cell-free DNA hypermethylation in colorectal cancer
Article Snippet: We distributed the first round reaction mix to individual 200 μl PCR tubes, which were incubated for five minutes at 37 °C, followed by 95 °C for five minutes, and cooled to room temperature. .. We added 10 μl of the first round PCR product to 710 μl preincubated reaction mix (37 °C for five minutes and 95 °C for 10 minutes) containing PCR buffer, 250 μM dNTP, 10 μM MgCl2 , 8 U Taq polymerase (Bioline® , [Taunton, MA, USA]), and 0.8 Uracil-DNA Glycosylase (Invitrogen® [Waltham, MA, USA]).

Article Title: The Use of Fluorescent Fragment Length Analysis (PCR-FFL) in the Direct Diagnosis and Identification of Cutaneous Leishmania Species
Article Snippet: Amplification reactions were performed in volumes of 50 μL containing 5 μL of 10X buffer (BIOTAQ DNA Polymerase, Bioline, London, UK), 1.5 mM MgCl2 , 0.2 mM dNTP, 0.2 μM of each primer and 1.5 units of Taq polymerase (BIOTAQ DNA Polymerase, Bioline, London, UK). .. We added 3 μL of isolated DNA to the mixture and incubated it in the thermal cycler (MJ Research PTC-200 DNA Engine, Alameda, CA) under the following conditions: for the ITS-1 PCR, we started with a denaturing step at 95°C for 2 min, followed by 35 cycles of denaturing for 20 sec at 95°C, annealing for 30 sec at 53°C, and extension for 1 min at 72°C, followed by a final extension at 72°C for 1 h. For the 7SL PCR, the denaturing step lasted for 5 min at 95°C, followed by 35 cycles of denaturing for 20 sec at 95°C, annealing for 30 sec at 65°C, and extension for 1 min at 72°C.

Expressing:

Article Title: Modification of a Rodent Hindlimb Model of Secondary Lymphedema: Surgical Radicality versus Radiotherapeutic Ablation
Article Snippet: Paragraph title: 2.5. In Vitro Lymphatic Differentiation of MDSCs and Lymphatic Marker Expression ... The denatured RNA was mixed with 4 μ L of 5x reaction buffer, 1 mM dNTP, 20 U of RNase inhibitor, and reverse transcriptase BioScript (Bioline) to make a final volume of 20 μ LcDNA.

Activated Clotting Time Assay:

Article Title: The Colletotrichum acutatum species complex
Article Snippet: .. The GAPDH, CHS-1, HIS3, ACT and TUB2 PCR mixture contained 1 μL 20x diluted genomic DNA, 0.2 μM of each primer, 1x PCR buffer (Bioline, Luckenwalde, Germany), 2 mM MgCl2 , 20 μM of each dNTP, 0.7 μL DMSO and 0.25 U Taq DNA polymerase (Bioline). .. Conditions for PCR of these genes constituted an initial denaturation step of 5 min at 94 °C, followed by 40 cycles of 30 s at 94 °C, 30 s at 52 °C and 30 s at 72 °C, and a final denaturation step of 7 min at 72 °C, while the ITS PCR was performed as described by Woudenberg et al. ( ).

Article Title: Comparison of the microbial population in rabbits and guinea pigs by next generation sequencing
Article Snippet: PCR conditions PCR was performed to amplify part of the V2-V3 region 16S rRNA gene of each sample using the reverse primer 355R ( 5’-CTG CTG CCT CCC GTA GGA GT-3’ ) in all reactions [ ] and the basal forward primer 27F ( 5’-CCA TCT CAT CCC TGC GTG TCT CCG ACT CAG-3’ ) linked to a reaction sample-specific “bar code” 10mer at the 5’ of the primer. .. Approximately 1ng of DNA was used per reaction in a reaction cocktail containing both primers (100nM each), 200μM of each dNTP, the manufacturer’s buffer (supplemented to 1.8mM MgCl2 ) and 1.25U FastStart high fidelity enzyme (Bioline) in a final reaction volume of 25μl.

Countercurrent Chromatography:

Article Title: Comparison of the microbial population in rabbits and guinea pigs by next generation sequencing
Article Snippet: PCR conditions PCR was performed to amplify part of the V2-V3 region 16S rRNA gene of each sample using the reverse primer 355R ( 5’-CTG CTG CCT CCC GTA GGA GT-3’ ) in all reactions [ ] and the basal forward primer 27F ( 5’-CCA TCT CAT CCC TGC GTG TCT CCG ACT CAG-3’ ) linked to a reaction sample-specific “bar code” 10mer at the 5’ of the primer. .. Approximately 1ng of DNA was used per reaction in a reaction cocktail containing both primers (100nM each), 200μM of each dNTP, the manufacturer’s buffer (supplemented to 1.8mM MgCl2 ) and 1.25U FastStart high fidelity enzyme (Bioline) in a final reaction volume of 25μl.

Ligation:

Article Title: Association of factor H autoantibodies with deletions of CFHR1, CFHR3, CFHR4, and with mutations in CFH, CFI, CD46, and C3 in patients with atypical hemolytic uremic syndrome
Article Snippet: CFHR1 and CFHR3 copy number was measured by the use of multiplex ligation-dependent probe amplification with a kit from MRC Holland (SALSA MLPA kit P236-A1 ARMD). .. PCRs were carried out in 25-μL volumes with the use of 150 ng of DNA and contained 0.5mM each dNTP, 6.7mM MgCl2 , 12.5pM of each primer, 1 U of a hot-start Taq polymerase (Immolase, Bioline) in a buffer of 16mM (NH4 )2 SO4 , 67mM Tris-HCI, and 0.01% Tween-20.

Reverse Transcription Polymerase Chain Reaction:

Article Title: Somatostatin and opioid receptors do not regulate proliferation or apoptosis of the human multiple myeloma U266 cells
Article Snippet: Paragraph title: RT-PCR ... PCRs were performed using 2 μL of cDNAs in the PCR buffer supplied with the Taq polymerase supplemented with 1.5 mM MgCl2 , 0.2 mM of dNTP, 2.5 units of Taq polymerase (Bioline), and 0.5 μM of each sense and antisense primer.

Article Title: Modification of a Rodent Hindlimb Model of Secondary Lymphedema: Surgical Radicality versus Radiotherapeutic Ablation
Article Snippet: Reverse transcriptase polymerase chain reaction (RT-PCR) was performed for lymphatic markers Prox-1, Flt-4, and podoplanin. .. The denatured RNA was mixed with 4 μ L of 5x reaction buffer, 1 mM dNTP, 20 U of RNase inhibitor, and reverse transcriptase BioScript (Bioline) to make a final volume of 20 μ LcDNA.

Generated:

Article Title: The Colletotrichum acutatum species complex
Article Snippet: The GAPDH, CHS-1, HIS3, ACT and TUB2 PCR mixture contained 1 μL 20x diluted genomic DNA, 0.2 μM of each primer, 1x PCR buffer (Bioline, Luckenwalde, Germany), 2 mM MgCl2 , 20 μM of each dNTP, 0.7 μL DMSO and 0.25 U Taq DNA polymerase (Bioline). .. The DNA sequences generated with forward and reverse primers were used to obtain consensus sequences using Bionumerics v. 4.60 (Applied Maths, St-Marthens-Lathem, Belgium), and the alignment assembled and manually adjusted using Sequence Alignment Editor v. 2.0a11 ( ).

Sequencing:

Article Title: The Colletotrichum acutatum species complex
Article Snippet: The GAPDH, CHS-1, HIS3, ACT and TUB2 PCR mixture contained 1 μL 20x diluted genomic DNA, 0.2 μM of each primer, 1x PCR buffer (Bioline, Luckenwalde, Germany), 2 mM MgCl2 , 20 μM of each dNTP, 0.7 μL DMSO and 0.25 U Taq DNA polymerase (Bioline). .. The DNA sequences generated with forward and reverse primers were used to obtain consensus sequences using Bionumerics v. 4.60 (Applied Maths, St-Marthens-Lathem, Belgium), and the alignment assembled and manually adjusted using Sequence Alignment Editor v. 2.0a11 ( ).

Article Title: Multiple Ethnic Origins of Mitochondrial DNA Lineages for the Population of Mauritius
Article Snippet: The PCR was carried out in 50-μl volumes, containing 1X Tris–HCl buffer, 200 μM of each dNTP, 2.5 mM MgCl2, 50 pmoles of each primer and 3 U of Taq polymerase (Bioline). .. For those haplotypes that could not be classified based solely on HVR sequence, several SNPs (1473, 1888, 3432, 10295, 10556, 12285, 12561, 14308, 14569, 15287, 15355, 15431, 15497 and 15968) were analysed by sequencing using PCR primer pairs and published protocols .

Immunofluorescence:

Article Title: Modification of a Rodent Hindlimb Model of Secondary Lymphedema: Surgical Radicality versus Radiotherapeutic Ablation
Article Snippet: For immunofluorescence (IF) staining, the respective fluorescein isothiocyanate-(FITC-labeled) secondary antibody was incubated for 3 h at 4°C and mounted with DAPI and the slides were observed under microscopy. .. The denatured RNA was mixed with 4 μ L of 5x reaction buffer, 1 mM dNTP, 20 U of RNase inhibitor, and reverse transcriptase BioScript (Bioline) to make a final volume of 20 μ LcDNA.

DNA Extraction:

Article Title: Multiple Ethnic Origins of Mitochondrial DNA Lineages for the Population of Mauritius
Article Snippet: DNA extraction was carried out using QIAmp DNA Mini Kit (QIAgen), following manufacturer recommendations. .. The PCR was carried out in 50-μl volumes, containing 1X Tris–HCl buffer, 200 μM of each dNTP, 2.5 mM MgCl2, 50 pmoles of each primer and 3 U of Taq polymerase (Bioline).

Marker:

Article Title: Somatostatin and opioid receptors do not regulate proliferation or apoptosis of the human multiple myeloma U266 cells
Article Snippet: PCRs were performed using 2 μL of cDNAs in the PCR buffer supplied with the Taq polymerase supplemented with 1.5 mM MgCl2 , 0.2 mM of dNTP, 2.5 units of Taq polymerase (Bioline), and 0.5 μM of each sense and antisense primer. .. PCR products were run on a 1.5% agarose or 2% NuSieve® agarose gel with a 100 bp marker (Invitrogen) and stained with ethidium bromide.

Article Title: Modification of a Rodent Hindlimb Model of Secondary Lymphedema: Surgical Radicality versus Radiotherapeutic Ablation
Article Snippet: Paragraph title: 2.5. In Vitro Lymphatic Differentiation of MDSCs and Lymphatic Marker Expression ... The denatured RNA was mixed with 4 μ L of 5x reaction buffer, 1 mM dNTP, 20 U of RNase inhibitor, and reverse transcriptase BioScript (Bioline) to make a final volume of 20 μ LcDNA.

Methylation:

Article Title: The prognostic efficacy of cell-free DNA hypermethylation in colorectal cancer
Article Snippet: For the second PCR reaction, we distributed 10 μl buffer containing 0.4 μM methylation specific inner primers and probes in 30 individual wells (one for each promoter region) in a 96 well PCR plate. .. We added 10 μl of the first round PCR product to 710 μl preincubated reaction mix (37 °C for five minutes and 95 °C for 10 minutes) containing PCR buffer, 250 μM dNTP, 10 μM MgCl2 , 8 U Taq polymerase (Bioline® , [Taunton, MA, USA]), and 0.8 Uracil-DNA Glycosylase (Invitrogen® [Waltham, MA, USA]).

Isolation:

Article Title: Somatostatin and opioid receptors do not regulate proliferation or apoptosis of the human multiple myeloma U266 cells
Article Snippet: RT-PCR Total RNAs were extracted using the RNAgents® Total RNA Isolation System (Promega) according to Chomczynski and Sacchi [ ]. cDNAs were synthesized from 2 μg of RNA in a buffer supplied with the reverse transcriptase (RT) (Promega) containing 900 μM dNTP (Amersham), 20 units RNAsine (Promega), 500 ng random primers (Promega) and 200 units of Moloney murine leukaemia virus RT in a final volume of 20 μL. .. PCRs were performed using 2 μL of cDNAs in the PCR buffer supplied with the Taq polymerase supplemented with 1.5 mM MgCl2 , 0.2 mM of dNTP, 2.5 units of Taq polymerase (Bioline), and 0.5 μM of each sense and antisense primer.

Article Title: The Use of Fluorescent Fragment Length Analysis (PCR-FFL) in the Direct Diagnosis and Identification of Cutaneous Leishmania Species
Article Snippet: Amplification reactions were performed in volumes of 50 μL containing 5 μL of 10X buffer (BIOTAQ DNA Polymerase, Bioline, London, UK), 1.5 mM MgCl2 , 0.2 mM dNTP, 0.2 μM of each primer and 1.5 units of Taq polymerase (BIOTAQ DNA Polymerase, Bioline, London, UK). .. We added 3 μL of isolated DNA to the mixture and incubated it in the thermal cycler (MJ Research PTC-200 DNA Engine, Alameda, CA) under the following conditions: for the ITS-1 PCR, we started with a denaturing step at 95°C for 2 min, followed by 35 cycles of denaturing for 20 sec at 95°C, annealing for 30 sec at 53°C, and extension for 1 min at 72°C, followed by a final extension at 72°C for 1 h. For the 7SL PCR, the denaturing step lasted for 5 min at 95°C, followed by 35 cycles of denaturing for 20 sec at 95°C, annealing for 30 sec at 65°C, and extension for 1 min at 72°C.

Size-exclusion Chromatography:

Article Title: Predominant Role of Host Genetics in Controlling the Composition of Gut Microbiota
Article Snippet: The PCR reaction mixture contained 10 mM Tris-HCl, 2 mM MgCl2 , 50 mM KCl, 200 µM of each dNTP, 10.0 pmol of each primer, 1U of Bio-X-Act Short DNA polymerase (Bioline, UK), and 100 ng of template DNA in a final volume of 50 µl. .. To reduce PCR bias caused by elevated number of PCR cycles , amplifications were performed using the following conditions: initial denaturation of template DNA at 94°C for 5 min, followed by 10 cycles of denaturation at 94°C for 30 sec, annealing at 57°C for 30 sec, and extension at 72°C for 2 min, with a final extension at 72°C for 10 min. PCR products were visualized on 1% agarose gel in TBE buffer stained with GelStar (Cambrex, UK).

Article Title: Assortative mating among Lake Malawi cichlid fish populations is not simply predictable from male nuptial colour
Article Snippet: .. PCR conditions consisted of 30 cycles of 30 sec at 95°, 60 sec at 55°, 60 sec at 72° in a 20 μl reaction volume containing 0.25 mM of each dNTP, 2 μl of 10× reaction buffer (Bioline, London, UK) 1.5 mM MgCl2 , 0.5 U of taq DNA polymerase (Bioline, London, UK), 25 ρmol of each primer and approximately 1 μg of template genomic DNA in a TGradient thermocycler (Biometra, Göttingen, Germany). .. PCR products were resolved on an ABI 310 (Applied Biosystems, Foster City, U.S.A.), with ROX-labelled internal size standards.

Article Title: Early infant diagnosis of HIV-1 infection in Luanda, Angola, using a new DNA PCR assay and dried blood spots
Article Snippet: The 25 μL reaction volume contained 1X NH4 buffer, 3 mmol/L MgCl2, 0.5 μmol/L of each dNTP, 0.3 μmol/L forward and reverse primers , 1U of Taq DNA polymerase (Bioline® Reagents Ltd, London, UK) plus 2.5 μL of DNA solution from the clinical or control samples (first-round PCR). .. Amplification cycling conditions were as follows: denaturation step of 94°C/3 min followed by 35 cycles of denaturation at 94°C/ 45 sec, annealing at 56°C/35 sec, extension at 72°C/ 1 min followed by a single final extension step at 72°C/ 15 min. Amplified products were visualized with green safe staining after electrophoresis in 2% agarose gel.

Article Title: The Use of Fluorescent Fragment Length Analysis (PCR-FFL) in the Direct Diagnosis and Identification of Cutaneous Leishmania Species
Article Snippet: Amplification reactions were performed in volumes of 50 μL containing 5 μL of 10X buffer (BIOTAQ DNA Polymerase, Bioline, London, UK), 1.5 mM MgCl2 , 0.2 mM dNTP, 0.2 μM of each primer and 1.5 units of Taq polymerase (BIOTAQ DNA Polymerase, Bioline, London, UK). .. We added 3 μL of isolated DNA to the mixture and incubated it in the thermal cycler (MJ Research PTC-200 DNA Engine, Alameda, CA) under the following conditions: for the ITS-1 PCR, we started with a denaturing step at 95°C for 2 min, followed by 35 cycles of denaturing for 20 sec at 95°C, annealing for 30 sec at 53°C, and extension for 1 min at 72°C, followed by a final extension at 72°C for 1 h. For the 7SL PCR, the denaturing step lasted for 5 min at 95°C, followed by 35 cycles of denaturing for 20 sec at 95°C, annealing for 30 sec at 65°C, and extension for 1 min at 72°C.

Labeling:

Article Title: Optimizing Production in the New Generation of Apricot Cultivars: Self-incompatibility, S-RNase Allele Identification, and Incompatibility Group Assignment
Article Snippet: S-RNase allele identification by PCR analysis Amplification reactions for the first intron region of the S-RNase gene were carried out with the combination of the fluorescently labeled forward primer SRc-F (5′-CTCGCTTTCCTTGTTCTTGC-3′) with the reverse primer SRc-R (5′-GGCCATTGTTGCACAAATTG-3′; Romero et al., ; Vilanova et al., ). .. PCR amplifications were carried out in 15 μl reaction volumes, containing 10x NH4 Reaction Buffer, 25 mM Cl2 Mg, 2.5 mM of each dNTP, 10 μM of each primer, 100 ng of genomic DNA and 0.5 U of BioTaq™ DNA polymerase (Bioline, London, UK).

Article Title: The Use of Fluorescent Fragment Length Analysis (PCR-FFL) in the Direct Diagnosis and Identification of Cutaneous Leishmania Species
Article Snippet: For the 7SL region, we used the primers TRY7SL.For1 (5′-TGCTCTGTAACCTTCGGGGGCT-3′) and TRY7SL.Rev1 (5′-GGCTGCTCCGTYNCCGGCCTGACCC-3′)., These primers were also fluorescently labeled with the same fluorochromes (6-FAM and VIC), respectively (Applied Biosystems). .. Amplification reactions were performed in volumes of 50 μL containing 5 μL of 10X buffer (BIOTAQ DNA Polymerase, Bioline, London, UK), 1.5 mM MgCl2 , 0.2 mM dNTP, 0.2 μM of each primer and 1.5 units of Taq polymerase (BIOTAQ DNA Polymerase, Bioline, London, UK).

Purification:

Article Title: Predominant Role of Host Genetics in Controlling the Composition of Gut Microbiota
Article Snippet: DNA was additionally purified by phenol-chloroform-isoamyl alcohol (25∶24∶1) extraction, ethanol precipitated, and dissolved in TE buffer. .. The PCR reaction mixture contained 10 mM Tris-HCl, 2 mM MgCl2 , 50 mM KCl, 200 µM of each dNTP, 10.0 pmol of each primer, 1U of Bio-X-Act Short DNA polymerase (Bioline, UK), and 100 ng of template DNA in a final volume of 50 µl.

Article Title: The prognostic efficacy of cell-free DNA hypermethylation in colorectal cancer
Article Snippet: Thereafter, we added 25 μl of purified deamination product to each tube, and performed the PCR reaction for 20 rounds (92 °C for 15 seconds, 55 °C for 30 seconds, and 72 °C for 30 seconds). .. We added 10 μl of the first round PCR product to 710 μl preincubated reaction mix (37 °C for five minutes and 95 °C for 10 minutes) containing PCR buffer, 250 μM dNTP, 10 μM MgCl2 , 8 U Taq polymerase (Bioline® , [Taunton, MA, USA]), and 0.8 Uracil-DNA Glycosylase (Invitrogen® [Waltham, MA, USA]).

Polymerase Chain Reaction:

Article Title: Predominant Role of Host Genetics in Controlling the Composition of Gut Microbiota
Article Snippet: .. The PCR reaction mixture contained 10 mM Tris-HCl, 2 mM MgCl2 , 50 mM KCl, 200 µM of each dNTP, 10.0 pmol of each primer, 1U of Bio-X-Act Short DNA polymerase (Bioline, UK), and 100 ng of template DNA in a final volume of 50 µl. .. To reduce PCR bias caused by elevated number of PCR cycles , amplifications were performed using the following conditions: initial denaturation of template DNA at 94°C for 5 min, followed by 10 cycles of denaturation at 94°C for 30 sec, annealing at 57°C for 30 sec, and extension at 72°C for 2 min, with a final extension at 72°C for 10 min. PCR products were visualized on 1% agarose gel in TBE buffer stained with GelStar (Cambrex, UK).

Article Title: The Colletotrichum acutatum species complex
Article Snippet: .. The GAPDH, CHS-1, HIS3, ACT and TUB2 PCR mixture contained 1 μL 20x diluted genomic DNA, 0.2 μM of each primer, 1x PCR buffer (Bioline, Luckenwalde, Germany), 2 mM MgCl2 , 20 μM of each dNTP, 0.7 μL DMSO and 0.25 U Taq DNA polymerase (Bioline). .. Conditions for PCR of these genes constituted an initial denaturation step of 5 min at 94 °C, followed by 40 cycles of 30 s at 94 °C, 30 s at 52 °C and 30 s at 72 °C, and a final denaturation step of 7 min at 72 °C, while the ITS PCR was performed as described by Woudenberg et al. ( ).

Article Title: Assortative mating among Lake Malawi cichlid fish populations is not simply predictable from male nuptial colour
Article Snippet: .. PCR conditions consisted of 30 cycles of 30 sec at 95°, 60 sec at 55°, 60 sec at 72° in a 20 μl reaction volume containing 0.25 mM of each dNTP, 2 μl of 10× reaction buffer (Bioline, London, UK) 1.5 mM MgCl2 , 0.5 U of taq DNA polymerase (Bioline, London, UK), 25 ρmol of each primer and approximately 1 μg of template genomic DNA in a TGradient thermocycler (Biometra, Göttingen, Germany). .. PCR products were resolved on an ABI 310 (Applied Biosystems, Foster City, U.S.A.), with ROX-labelled internal size standards.

Article Title: Early infant diagnosis of HIV-1 infection in Luanda, Angola, using a new DNA PCR assay and dried blood spots
Article Snippet: .. The 25 μL reaction volume contained 1X NH4 buffer, 3 mmol/L MgCl2, 0.5 μmol/L of each dNTP, 0.3 μmol/L forward and reverse primers , 1U of Taq DNA polymerase (Bioline® Reagents Ltd, London, UK) plus 2.5 μL of DNA solution from the clinical or control samples (first-round PCR). .. For the second-round PCR reaction we used 2.5ul of the first-round PCR reaction.

Article Title: Comparison of the microbial population in rabbits and guinea pigs by next generation sequencing
Article Snippet: Paragraph title: PCR conditions ... Approximately 1ng of DNA was used per reaction in a reaction cocktail containing both primers (100nM each), 200μM of each dNTP, the manufacturer’s buffer (supplemented to 1.8mM MgCl2 ) and 1.25U FastStart high fidelity enzyme (Bioline) in a final reaction volume of 25μl.

Article Title: Optimizing Production in the New Generation of Apricot Cultivars: Self-incompatibility, S-RNase Allele Identification, and Incompatibility Group Assignment
Article Snippet: .. PCR amplifications were carried out in 15 μl reaction volumes, containing 10x NH4 Reaction Buffer, 25 mM Cl2 Mg, 2.5 mM of each dNTP, 10 μM of each primer, 100 ng of genomic DNA and 0.5 U of BioTaq™ DNA polymerase (Bioline, London, UK). .. The sizes of the products obtained by PCR were analyzed in a CEQ™ 8000 capillary electrophoresis DNA analysis system (Beckman Coulter, Fullerton, CA, USA) and compared and classified according to Vilanova et al. ( ) and Kodad et al. ( ).

Article Title: Somatostatin and opioid receptors do not regulate proliferation or apoptosis of the human multiple myeloma U266 cells
Article Snippet: .. PCRs were performed using 2 μL of cDNAs in the PCR buffer supplied with the Taq polymerase supplemented with 1.5 mM MgCl2 , 0.2 mM of dNTP, 2.5 units of Taq polymerase (Bioline), and 0.5 μM of each sense and antisense primer. ..

Article Title: Multiple Ethnic Origins of Mitochondrial DNA Lineages for the Population of Mauritius
Article Snippet: .. The PCR was carried out in 50-μl volumes, containing 1X Tris–HCl buffer, 200 μM of each dNTP, 2.5 mM MgCl2, 50 pmoles of each primer and 3 U of Taq polymerase (Bioline). .. The amplification was carried out in an Applied Biosystems 2720 Thermal Cycler with the following conditions: 30 amplification cycles with denaturation at 94°C for 30 s, annealing at 55°C for 30 s, and extension at 72°C for 90 s. HVR haplotypes were classified into haplogroups based on the mtDNA tree (Build 15) .

Article Title: Modification of a Rodent Hindlimb Model of Secondary Lymphedema: Surgical Radicality versus Radiotherapeutic Ablation
Article Snippet: Reverse transcriptase polymerase chain reaction (RT-PCR) was performed for lymphatic markers Prox-1, Flt-4, and podoplanin. .. The denatured RNA was mixed with 4 μ L of 5x reaction buffer, 1 mM dNTP, 20 U of RNase inhibitor, and reverse transcriptase BioScript (Bioline) to make a final volume of 20 μ LcDNA.

Article Title: The prognostic efficacy of cell-free DNA hypermethylation in colorectal cancer
Article Snippet: .. We added 10 μl of the first round PCR product to 710 μl preincubated reaction mix (37 °C for five minutes and 95 °C for 10 minutes) containing PCR buffer, 250 μM dNTP, 10 μM MgCl2 , 8 U Taq polymerase (Bioline® , [Taunton, MA, USA]), and 0.8 Uracil-DNA Glycosylase (Invitrogen® [Waltham, MA, USA]). ..

Article Title: The Use of Fluorescent Fragment Length Analysis (PCR-FFL) in the Direct Diagnosis and Identification of Cutaneous Leishmania Species
Article Snippet: Paragraph title: PCR amplification. ... Amplification reactions were performed in volumes of 50 μL containing 5 μL of 10X buffer (BIOTAQ DNA Polymerase, Bioline, London, UK), 1.5 mM MgCl2 , 0.2 mM dNTP, 0.2 μM of each primer and 1.5 units of Taq polymerase (BIOTAQ DNA Polymerase, Bioline, London, UK).

Article Title: Association of factor H autoantibodies with deletions of CFHR1, CFHR3, CFHR4, and with mutations in CFH, CFI, CD46, and C3 in patients with atypical hemolytic uremic syndrome
Article Snippet: CFHR4 copy number was measured by the use of a multiplex polymerase chain reaction (PCR) assay designed to amplify CFHR4 exon 2 and intron 1. .. PCRs were carried out in 25-μL volumes with the use of 150 ng of DNA and contained 0.5mM each dNTP, 6.7mM MgCl2 , 12.5pM of each primer, 1 U of a hot-start Taq polymerase (Immolase, Bioline) in a buffer of 16mM (NH4 )2 SO4 , 67mM Tris-HCI, and 0.01% Tween-20.

Microscopy:

Article Title: Modification of a Rodent Hindlimb Model of Secondary Lymphedema: Surgical Radicality versus Radiotherapeutic Ablation
Article Snippet: For immunofluorescence (IF) staining, the respective fluorescein isothiocyanate-(FITC-labeled) secondary antibody was incubated for 3 h at 4°C and mounted with DAPI and the slides were observed under microscopy. .. The denatured RNA was mixed with 4 μ L of 5x reaction buffer, 1 mM dNTP, 20 U of RNase inhibitor, and reverse transcriptase BioScript (Bioline) to make a final volume of 20 μ LcDNA.

Nested PCR:

Article Title: Early infant diagnosis of HIV-1 infection in Luanda, Angola, using a new DNA PCR assay and dried blood spots
Article Snippet: PCR amplification of proviral HIV-1 DNA A nested PCR was used to amplify a 194 bp fragment of the IN gene. .. The 25 μL reaction volume contained 1X NH4 buffer, 3 mmol/L MgCl2, 0.5 μmol/L of each dNTP, 0.3 μmol/L forward and reverse primers , 1U of Taq DNA polymerase (Bioline® Reagents Ltd, London, UK) plus 2.5 μL of DNA solution from the clinical or control samples (first-round PCR).

Multiplex Ligation-dependent Probe Amplification:

Article Title: Association of factor H autoantibodies with deletions of CFHR1, CFHR3, CFHR4, and with mutations in CFH, CFI, CD46, and C3 in patients with atypical hemolytic uremic syndrome
Article Snippet: CFHR1 and CFHR3 copy number was measured by the use of multiplex ligation-dependent probe amplification with a kit from MRC Holland (SALSA MLPA kit P236-A1 ARMD). .. PCRs were carried out in 25-μL volumes with the use of 150 ng of DNA and contained 0.5mM each dNTP, 6.7mM MgCl2 , 12.5pM of each primer, 1 U of a hot-start Taq polymerase (Immolase, Bioline) in a buffer of 16mM (NH4 )2 SO4 , 67mM Tris-HCI, and 0.01% Tween-20.

Chloramphenicol Acetyltransferase Assay:

Article Title: Comparison of the microbial population in rabbits and guinea pigs by next generation sequencing
Article Snippet: PCR conditions PCR was performed to amplify part of the V2-V3 region 16S rRNA gene of each sample using the reverse primer 355R ( 5’-CTG CTG CCT CCC GTA GGA GT-3’ ) in all reactions [ ] and the basal forward primer 27F ( 5’-CCA TCT CAT CCC TGC GTG TCT CCG ACT CAG-3’ ) linked to a reaction sample-specific “bar code” 10mer at the 5’ of the primer. .. Approximately 1ng of DNA was used per reaction in a reaction cocktail containing both primers (100nM each), 200μM of each dNTP, the manufacturer’s buffer (supplemented to 1.8mM MgCl2 ) and 1.25U FastStart high fidelity enzyme (Bioline) in a final reaction volume of 25μl.

SYBR Green Assay:

Article Title: Optimizing Production in the New Generation of Apricot Cultivars: Self-incompatibility, S-RNase Allele Identification, and Incompatibility Group Assignment
Article Snippet: PCR amplifications were carried out in 15 μl reaction volumes, containing 10x NH4 Reaction Buffer, 25 mM Cl2 Mg, 2.5 mM of each dNTP, 10 μM of each primer, 100 ng of genomic DNA and 0.5 U of BioTaq™ DNA polymerase (Bioline, London, UK). .. Amplified fragments of the second intron were separated on 1% (w/v) agarose gels and DNA bands were visualized using the nucleic acid stain SYBR Green (Thermo).

Multiplex Assay:

Article Title: Association of factor H autoantibodies with deletions of CFHR1, CFHR3, CFHR4, and with mutations in CFH, CFI, CD46, and C3 in patients with atypical hemolytic uremic syndrome
Article Snippet: CFHR1 copy number was measured by the use of the same multiplex assay designed to amplify CFHR1 introns 3 and 5. .. PCRs were carried out in 25-μL volumes with the use of 150 ng of DNA and contained 0.5mM each dNTP, 6.7mM MgCl2 , 12.5pM of each primer, 1 U of a hot-start Taq polymerase (Immolase, Bioline) in a buffer of 16mM (NH4 )2 SO4 , 67mM Tris-HCI, and 0.01% Tween-20.

Agarose Gel Electrophoresis:

Article Title: Predominant Role of Host Genetics in Controlling the Composition of Gut Microbiota
Article Snippet: The PCR reaction mixture contained 10 mM Tris-HCl, 2 mM MgCl2 , 50 mM KCl, 200 µM of each dNTP, 10.0 pmol of each primer, 1U of Bio-X-Act Short DNA polymerase (Bioline, UK), and 100 ng of template DNA in a final volume of 50 µl. .. To reduce PCR bias caused by elevated number of PCR cycles , amplifications were performed using the following conditions: initial denaturation of template DNA at 94°C for 5 min, followed by 10 cycles of denaturation at 94°C for 30 sec, annealing at 57°C for 30 sec, and extension at 72°C for 2 min, with a final extension at 72°C for 10 min. PCR products were visualized on 1% agarose gel in TBE buffer stained with GelStar (Cambrex, UK).

Article Title: Early infant diagnosis of HIV-1 infection in Luanda, Angola, using a new DNA PCR assay and dried blood spots
Article Snippet: The 25 μL reaction volume contained 1X NH4 buffer, 3 mmol/L MgCl2, 0.5 μmol/L of each dNTP, 0.3 μmol/L forward and reverse primers , 1U of Taq DNA polymerase (Bioline® Reagents Ltd, London, UK) plus 2.5 μL of DNA solution from the clinical or control samples (first-round PCR). .. Amplification cycling conditions were as follows: denaturation step of 94°C/3 min followed by 35 cycles of denaturation at 94°C/ 45 sec, annealing at 56°C/35 sec, extension at 72°C/ 1 min followed by a single final extension step at 72°C/ 15 min. Amplified products were visualized with green safe staining after electrophoresis in 2% agarose gel.

Article Title: Somatostatin and opioid receptors do not regulate proliferation or apoptosis of the human multiple myeloma U266 cells
Article Snippet: PCRs were performed using 2 μL of cDNAs in the PCR buffer supplied with the Taq polymerase supplemented with 1.5 mM MgCl2 , 0.2 mM of dNTP, 2.5 units of Taq polymerase (Bioline), and 0.5 μM of each sense and antisense primer. .. PCR products were run on a 1.5% agarose or 2% NuSieve® agarose gel with a 100 bp marker (Invitrogen) and stained with ethidium bromide.

In Vitro:

Article Title: Modification of a Rodent Hindlimb Model of Secondary Lymphedema: Surgical Radicality versus Radiotherapeutic Ablation
Article Snippet: Paragraph title: 2.5. In Vitro Lymphatic Differentiation of MDSCs and Lymphatic Marker Expression ... The denatured RNA was mixed with 4 μ L of 5x reaction buffer, 1 mM dNTP, 20 U of RNase inhibitor, and reverse transcriptase BioScript (Bioline) to make a final volume of 20 μ LcDNA.

Electrophoresis:

Article Title: Early infant diagnosis of HIV-1 infection in Luanda, Angola, using a new DNA PCR assay and dried blood spots
Article Snippet: The 25 μL reaction volume contained 1X NH4 buffer, 3 mmol/L MgCl2, 0.5 μmol/L of each dNTP, 0.3 μmol/L forward and reverse primers , 1U of Taq DNA polymerase (Bioline® Reagents Ltd, London, UK) plus 2.5 μL of DNA solution from the clinical or control samples (first-round PCR). .. Amplification cycling conditions were as follows: denaturation step of 94°C/3 min followed by 35 cycles of denaturation at 94°C/ 45 sec, annealing at 56°C/35 sec, extension at 72°C/ 1 min followed by a single final extension step at 72°C/ 15 min. Amplified products were visualized with green safe staining after electrophoresis in 2% agarose gel.

Article Title: Comparison of the microbial population in rabbits and guinea pigs by next generation sequencing
Article Snippet: Approximately 1ng of DNA was used per reaction in a reaction cocktail containing both primers (100nM each), 200μM of each dNTP, the manufacturer’s buffer (supplemented to 1.8mM MgCl2 ) and 1.25U FastStart high fidelity enzyme (Bioline) in a final reaction volume of 25μl. .. The presence of amplicons was verified at the end of the PCR stage by electrophoresis.

Article Title: Optimizing Production in the New Generation of Apricot Cultivars: Self-incompatibility, S-RNase Allele Identification, and Incompatibility Group Assignment
Article Snippet: PCR amplifications were carried out in 15 μl reaction volumes, containing 10x NH4 Reaction Buffer, 25 mM Cl2 Mg, 2.5 mM of each dNTP, 10 μM of each primer, 100 ng of genomic DNA and 0.5 U of BioTaq™ DNA polymerase (Bioline, London, UK). .. The sizes of the products obtained by PCR were analyzed in a CEQ™ 8000 capillary electrophoresis DNA analysis system (Beckman Coulter, Fullerton, CA, USA) and compared and classified according to Vilanova et al. ( ) and Kodad et al. ( ).

Article Title: Association of factor H autoantibodies with deletions of CFHR1, CFHR3, CFHR4, and with mutations in CFH, CFI, CD46, and C3 in patients with atypical hemolytic uremic syndrome
Article Snippet: PCRs were carried out in 25-μL volumes with the use of 150 ng of DNA and contained 0.5mM each dNTP, 6.7mM MgCl2 , 12.5pM of each primer, 1 U of a hot-start Taq polymerase (Immolase, Bioline) in a buffer of 16mM (NH4 )2 SO4 , 67mM Tris-HCI, and 0.01% Tween-20. .. PCRs were carried out in 25-μL volumes with the use of 150 ng of DNA and contained 0.5mM each dNTP, 6.7mM MgCl2 , 12.5pM of each primer, 1 U of a hot-start Taq polymerase (Immolase, Bioline) in a buffer of 16mM (NH4 )2 SO4 , 67mM Tris-HCI, and 0.01% Tween-20.

Ethanol Precipitation:

Article Title: Predominant Role of Host Genetics in Controlling the Composition of Gut Microbiota
Article Snippet: The PCR reaction mixture contained 10 mM Tris-HCl, 2 mM MgCl2 , 50 mM KCl, 200 µM of each dNTP, 10.0 pmol of each primer, 1U of Bio-X-Act Short DNA polymerase (Bioline, UK), and 100 ng of template DNA in a final volume of 50 µl. .. This was followed by ethanol precipitation and the final pellets were suspended in 5 µl of TE buffer (pH 8.0).

Spectrophotometry:

Article Title: Modification of a Rodent Hindlimb Model of Secondary Lymphedema: Surgical Radicality versus Radiotherapeutic Ablation
Article Snippet: Total RNA from the respective cells were extracted using the RNeasy Mini Kit (Qiagen) according to the manufacturer's instructions, and the quantity and quality of the extracted RNA were determined by absorbance at 260 nm and 280 nm wavelengths using the SmartSpec Plus Spectrophotometer (Bio-Rad). .. The denatured RNA was mixed with 4 μ L of 5x reaction buffer, 1 mM dNTP, 20 U of RNase inhibitor, and reverse transcriptase BioScript (Bioline) to make a final volume of 20 μ LcDNA.

CTG Assay:

Article Title: Comparison of the microbial population in rabbits and guinea pigs by next generation sequencing
Article Snippet: PCR conditions PCR was performed to amplify part of the V2-V3 region 16S rRNA gene of each sample using the reverse primer 355R ( 5’-CTG CTG CCT CCC GTA GGA GT-3’ ) in all reactions [ ] and the basal forward primer 27F ( 5’-CCA TCT CAT CCC TGC GTG TCT CCG ACT CAG-3’ ) linked to a reaction sample-specific “bar code” 10mer at the 5’ of the primer. .. Approximately 1ng of DNA was used per reaction in a reaction cocktail containing both primers (100nM each), 200μM of each dNTP, the manufacturer’s buffer (supplemented to 1.8mM MgCl2 ) and 1.25U FastStart high fidelity enzyme (Bioline) in a final reaction volume of 25μl.

Staining:

Article Title: Predominant Role of Host Genetics in Controlling the Composition of Gut Microbiota
Article Snippet: The PCR reaction mixture contained 10 mM Tris-HCl, 2 mM MgCl2 , 50 mM KCl, 200 µM of each dNTP, 10.0 pmol of each primer, 1U of Bio-X-Act Short DNA polymerase (Bioline, UK), and 100 ng of template DNA in a final volume of 50 µl. .. To reduce PCR bias caused by elevated number of PCR cycles , amplifications were performed using the following conditions: initial denaturation of template DNA at 94°C for 5 min, followed by 10 cycles of denaturation at 94°C for 30 sec, annealing at 57°C for 30 sec, and extension at 72°C for 2 min, with a final extension at 72°C for 10 min. PCR products were visualized on 1% agarose gel in TBE buffer stained with GelStar (Cambrex, UK).

Article Title: Early infant diagnosis of HIV-1 infection in Luanda, Angola, using a new DNA PCR assay and dried blood spots
Article Snippet: The 25 μL reaction volume contained 1X NH4 buffer, 3 mmol/L MgCl2, 0.5 μmol/L of each dNTP, 0.3 μmol/L forward and reverse primers , 1U of Taq DNA polymerase (Bioline® Reagents Ltd, London, UK) plus 2.5 μL of DNA solution from the clinical or control samples (first-round PCR). .. Amplification cycling conditions were as follows: denaturation step of 94°C/3 min followed by 35 cycles of denaturation at 94°C/ 45 sec, annealing at 56°C/35 sec, extension at 72°C/ 1 min followed by a single final extension step at 72°C/ 15 min. Amplified products were visualized with green safe staining after electrophoresis in 2% agarose gel.

Article Title: Optimizing Production in the New Generation of Apricot Cultivars: Self-incompatibility, S-RNase Allele Identification, and Incompatibility Group Assignment
Article Snippet: PCR amplifications were carried out in 15 μl reaction volumes, containing 10x NH4 Reaction Buffer, 25 mM Cl2 Mg, 2.5 mM of each dNTP, 10 μM of each primer, 100 ng of genomic DNA and 0.5 U of BioTaq™ DNA polymerase (Bioline, London, UK). .. Amplified fragments of the second intron were separated on 1% (w/v) agarose gels and DNA bands were visualized using the nucleic acid stain SYBR Green (Thermo).

Article Title: Somatostatin and opioid receptors do not regulate proliferation or apoptosis of the human multiple myeloma U266 cells
Article Snippet: PCRs were performed using 2 μL of cDNAs in the PCR buffer supplied with the Taq polymerase supplemented with 1.5 mM MgCl2 , 0.2 mM of dNTP, 2.5 units of Taq polymerase (Bioline), and 0.5 μM of each sense and antisense primer. .. PCR products were run on a 1.5% agarose or 2% NuSieve® agarose gel with a 100 bp marker (Invitrogen) and stained with ethidium bromide.

Article Title: Modification of a Rodent Hindlimb Model of Secondary Lymphedema: Surgical Radicality versus Radiotherapeutic Ablation
Article Snippet: For immunofluorescence (IF) staining, the respective fluorescein isothiocyanate-(FITC-labeled) secondary antibody was incubated for 3 h at 4°C and mounted with DAPI and the slides were observed under microscopy. .. The denatured RNA was mixed with 4 μ L of 5x reaction buffer, 1 mM dNTP, 20 U of RNase inhibitor, and reverse transcriptase BioScript (Bioline) to make a final volume of 20 μ LcDNA.