Polymerase Chain Reaction:Article Title: Improvement of Cryopreservation and Production of Attenuated Babesia Parasites to Prevent Bovine Babesiosis.
Article Snippet: .. Both single and nPCR protocols were carried out in a final volume of 25 μL; the mixture included 12.5 μL of Go 2x Taq Green Master Mix (400 μM of dNTP, 3 μM MgCl2, and 1.5 U Taq DNA polymerase in 2× buffer, pH 8.5; PROMEGA, Madison, WI, USA), 1 μL of each of the forward and reverse primers (10 pmol), and 80–120 ng of genomic DNA as the template for a single PCR reaction and 2 μL of the generated amplicon as the template for nPCR. ..
Article Title: Incipient Range Expansion of Green Turtles in the Mediterranean.
Article Snippet: .. We performed the polymerase chain reaction (PCR) in a final volume of 15 μL containing 2 μL of DNA, 5.08 μL of deionised water, 3 μL of PCR buffer 5× (GoTaq Promega), 1.8 μL of dNTPs (1 mm), 0.6 μL of MgCl2 (25 mm), 1.8 μL of bovine serum albumin, 0.3 μL of each primer (10 μm) and 0.12 μL of GoTaq G2 Flexi DNA Polymerase (Promega, 5 U/μL). .. Next, we purified 3 μL of the product with 2 μL of ExoSAP (0.4 U of EXO and 0.4 U of TSAP) through a single cycle of incubation at 37°C for 15 min followed by 80°C for another 15 min. Then, we introduced 1 μL (5 μm) of forward primer (LCM15382) and we dried at 80°C for 30 min, preparing the mtDNA amplicons for sequencing on an ABI 3730 automated DNA analyser (Applied Biosystems) at the Scientific and Technical Services of the University of Barcelona.
Article Title: Incipient Range Expansion of Green Turtles in the Mediterranean
Article Snippet: .. We performed the polymerase chain reaction (PCR) in a final volume of 15 μL containing 2 μL of DNA, 5.08 μL of deionised water, 3 μL of PCR buffer 5× (GoTaq Promega), 1.8 μL of dNTPs (1 mm), 0.6 μL of MgCl 2 (25 mm), 1.8 μL of bovine serum albumin, 0.3 μL of each primer (10 μm) and 0.12 μL of GoTaq G2 Flexi DNA Polymerase (Promega, 5 U/μL). .. Next, we purified 3 μL of the product with 2 μL of ExoSAP (0.4 U of EXO and 0.4 U of TSAP) through a single cycle of incubation at 37°C for 15 min followed by 80°C for another 15 min. Then, we introduced 1 μL (5 μm) of forward primer (LCM15382) and we dried at 80°C for 30 min, preparing the mtDNA amplicons for sequencing on an ABI 3730 automated DNA analyser (Applied Biosystems) at the Scientific and Technical Services of the University of Barcelona.
Article Title: Matrigel inhibits elongation and drives endoderm differentiation in aggregates of mouse embryonic stem cells.
Article Snippet: A minimum of 1 μg of RNA was combined with 1 μL of Oligo (dT)15 primer (C110B; Promega) and nuclease-free water to make up a total of 5 μL and incubated at 70 °C and 4 °C for 5min each. .. PCR master mix made up of 6.1 μL nuclease-free water, 4 μL 5× reaction buffer (M289A; Promega), 2.4 μL MgCl2 (A351H; Promega), 1 dNTP mix (C114B; Promega), 0.5 μL Recombinant RNasin® Ribonuclease Inhibitor (N251A; Promega), and 1 μL Reverse Transcriptase (M314A; Promega), was added to each sample. .. Thermal cycling was as follows: 25 °C for 5min, 42 °C for 60min and 70 °C for 15min. qPCR The StepOnePlusTM Real-Time PCR System was used for quantitative PCR with SYBR green PCR Master-Mix (4368708; Thermo-Fisher Scientific).
Generated:Article Title: Improvement of Cryopreservation and Production of Attenuated Babesia Parasites to Prevent Bovine Babesiosis.
Article Snippet: .. Both single and nPCR protocols were carried out in a final volume of 25 μL; the mixture included 12.5 μL of Go 2x Taq Green Master Mix (400 μM of dNTP, 3 μM MgCl2, and 1.5 U Taq DNA polymerase in 2× buffer, pH 8.5; PROMEGA, Madison, WI, USA), 1 μL of each of the forward and reverse primers (10 pmol), and 80–120 ng of genomic DNA as the template for a single PCR reaction and 2 μL of the generated amplicon as the template for nPCR. ..
Amplification:Article Title: Improvement of Cryopreservation and Production of Attenuated Babesia Parasites to Prevent Bovine Babesiosis.
Article Snippet: .. Both single and nPCR protocols were carried out in a final volume of 25 μL; the mixture included 12.5 μL of Go 2x Taq Green Master Mix (400 μM of dNTP, 3 μM MgCl2, and 1.5 U Taq DNA polymerase in 2× buffer, pH 8.5; PROMEGA, Madison, WI, USA), 1 μL of each of the forward and reverse primers (10 pmol), and 80–120 ng of genomic DNA as the template for a single PCR reaction and 2 μL of the generated amplicon as the template for nPCR. ..
Recombinant:Article Title: Matrigel inhibits elongation and drives endoderm differentiation in aggregates of mouse embryonic stem cells.
Article Snippet: A minimum of 1 μg of RNA was combined with 1 μL of Oligo (dT)15 primer (C110B; Promega) and nuclease-free water to make up a total of 5 μL and incubated at 70 °C and 4 °C for 5min each. .. PCR master mix made up of 6.1 μL nuclease-free water, 4 μL 5× reaction buffer (M289A; Promega), 2.4 μL MgCl2 (A351H; Promega), 1 dNTP mix (C114B; Promega), 0.5 μL Recombinant RNasin® Ribonuclease Inhibitor (N251A; Promega), and 1 μL Reverse Transcriptase (M314A; Promega), was added to each sample. .. Thermal cycling was as follows: 25 °C for 5min, 42 °C for 60min and 70 °C for 15min. qPCR The StepOnePlusTM Real-Time PCR System was used for quantitative PCR with SYBR green PCR Master-Mix (4368708; Thermo-Fisher Scientific).
Reverse Transcription:Article Title: Matrigel inhibits elongation and drives endoderm differentiation in aggregates of mouse embryonic stem cells.
Article Snippet: A minimum of 1 μg of RNA was combined with 1 μL of Oligo (dT)15 primer (C110B; Promega) and nuclease-free water to make up a total of 5 μL and incubated at 70 °C and 4 °C for 5min each. .. PCR master mix made up of 6.1 μL nuclease-free water, 4 μL 5× reaction buffer (M289A; Promega), 2.4 μL MgCl2 (A351H; Promega), 1 dNTP mix (C114B; Promega), 0.5 μL Recombinant RNasin® Ribonuclease Inhibitor (N251A; Promega), and 1 μL Reverse Transcriptase (M314A; Promega), was added to each sample. .. Thermal cycling was as follows: 25 °C for 5min, 42 °C for 60min and 70 °C for 15min. qPCR The StepOnePlusTM Real-Time PCR System was used for quantitative PCR with SYBR green PCR Master-Mix (4368708; Thermo-Fisher Scientific).
other:Article Title: EIF3D safeguards the homeostasis of key signaling pathways in human primed pluripotency
Article Snippet: P3 Primary Cell 4D-Nucleofector X Kit S Lonza Cat# V4XP-3032 TransIT-Lenti Transfection Reagent Mirus Cat# MIR6600 Lenti-X qRT-PCR Titration Kit Takara Cat# 631235 Lipofectamine RNAi Max reagent Thermo Fisher Scientific Cat# 13778150 Click-iT EdU Alexa Fluor 647 Flow Cytometry Assay Kit Thermo Fisher Scientific Cat# C10424 QIAzol lysis reagent QIAGEN Cat# 79306 RIPA Buffer Sigma-Aldrich Cat# R0278 Passive 5× Lysis Buffer Promega Cat# E1941 TRIZOL LS reagent Thermo Fisher Scientific Cat# 10296028 Direct-zol RNA Miniprep kit Zymo Research Cat# R2052 Direct-zol RNA Microprep kit Zymo Research Cat# R2062 ReverTra Ace qPCR RT Kit TOYOBO Cat# FSQ-101 THUNDERBIRD Next SYBR qPCR Mix TOYOBO Cat# QPX-201 TaqMan Universal Master Mix II, no UNG Thermo Fisher Scientific Cat# 4440040 NucleoSpin Blood XL Takara U0950B QIAamp DNA Blood Midi Kit QIAGEN Cat# 51185 QIAquick Gel Extraction Kit QIAGEN Cat# 28706 SbfI-HF New England Biolabs Cat# R3642 KOD One Master Mix TOYOBO Cat# KMM-101 NEBuilder HiFi DNA Assembly Master Mix New England Biolabs Cat# E2621 riboPOOLs for Ribo-seq (Homo sapiens) siTOOLs Biotech Cat# 042 riboPOOLs (Homo sapiens) siTOOLs Biotech Cat# 054 xGen UDI-UMI Adapters Integrated DNA Technologies Cat# 10005903 ProtoScript II New England Biolabs Cat# M0368L CircLigaseII ssDNA ligase Epicentre Cat# CL9025K Turbo DNase Thermo Fisher Scientific Cat# AM2238 RNase I Epicentre Cat# N6901K SUPERase-In Thermo Fisher Scientific Cat# AM2696 Phusion polymerase New England Biolabs Cat# M0530S Dr.
Article Title: Identifying biomarkers of sheep welfare using a metabolic discrepancy model.
Article Snippet: Each reverse transcription reaction contained 0.5 μL of Random Primers (Cat.C1181, Promega, Aus); 5.5 μL total RNA after DNA digestion; and 8 μL RNase-free water and was incubated at 70 °C for 5 min and chilled at 4 °C for 5 min. Then 5 μL of M-MLV RT 5X Reaction Buffer (Cat.M1705, Promega, Aus); 1.3 μL of dNTP Mix (Cat.U1511, Promega, Aus); 1 μL of M-MLV RT (H-) Point Mutant (Cat.M3682, Promega, Aus); and 3.7 μL of RNase-free water was added to each reaction and the reverse transcription protocol was performed at 25 °C for 10 min, 55 °C for 50 min, and 70 °C for 15 min.
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