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Promega wizard dna clean up system
Wizard Dna Clean Up System, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 289 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 99 stars, based on 289 article reviews
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wizard dna clean up system - by Bioz Stars, 2020-04
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Clone Assay:

Article Title: Upregulation of TRAG3 gene in urothelial carcinoma of the bladder
Article Snippet: Modified samples were purified using Wizard DNA Clean-Up System desalting columns (Promega) followed by ethanol precipitation. .. The PCR products were cloned into TA cloning vector pCR 2.1-TOPO (Invitrogen).

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: Next, targeting ORF3 was amplified for 45 cycles at 94°C for 15 s and 60°C for 60 s. In addition, to generate the internal control RNA, a 174-bp RT-PCR product was amplified using the primers Taqman-T7f and Taqman-R ( ) and then cloned into the pMD19-T vector (TaKaRa Biotech Corporation, Dalian, China). .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol.

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: In addition, to generate the internal control RNA, a 174-bp RT-PCR product was amplified using the primers Taqman-T7f and Taqman-R ( ) and then cloned into the pMD19-T vector (TaKaRa Biotech Corporation, Dalian, China). .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol.

Amplification:

Article Title: Upregulation of TRAG3 gene in urothelial carcinoma of the bladder
Article Snippet: Modified samples were purified using Wizard DNA Clean-Up System desalting columns (Promega) followed by ethanol precipitation. .. Bisulfite-modified DNA (100 ng) was PCR amplified (35 cycles: 95 °C, 30 sec; 54 °C, 30 sec; 72 °C, 1 min) with primer sets described previously in a 25-µl reaction mixture.

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: Next, targeting ORF3 was amplified for 45 cycles at 94°C for 15 s and 60°C for 60 s. In addition, to generate the internal control RNA, a 174-bp RT-PCR product was amplified using the primers Taqman-T7f and Taqman-R ( ) and then cloned into the pMD19-T vector (TaKaRa Biotech Corporation, Dalian, China). .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol.

Article Title: Applying DNA barcoding to red macroalgae: a preliminary appraisal holds promise for future applications
Article Snippet: DNA was extracted with a protocol modified from (instead of the final agarose gel cleaning procedure, the DNA was purified with the Wizard® DNA Clean-Up System, Promega Corp., Madison, WI). .. The PCR amplification profile followed , but using an annealing temperature of 50 °C.

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: In addition, to generate the internal control RNA, a 174-bp RT-PCR product was amplified using the primers Taqman-T7f and Taqman-R ( ) and then cloned into the pMD19-T vector (TaKaRa Biotech Corporation, Dalian, China). .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol.

Article Title: Stool-based DNA testing, a new noninvasive method for colorectal cancer screening, the first report from Iran
Article Snippet: Modified DNA was purified using a Promega Wizard DNA Clean-Up System, according to the manufacturer’s instructions and then was stored at -20°C until it was used for PCR. .. Modified DNA was amplified using primers specific for methylated and unmethylated p 16 sequences as previously described[ ].

Synthesized:

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: The cDNA from RNA was synthesized via reverse transcription for 30 min at 50°C, followed by an PCR initial activation step for 15 min at 95°C. .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol.

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: The cDNA from RNA was synthesized via reverse transcription for 30 min at 50°C, followed by an PCR initial activation step for 15 min at 95°C. .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol.

Neutralization:

Article Title: Activation of PI3K/mTOR pathway occurs in most adult low-grade gliomas and predicts patient survival
Article Snippet: Bisulfite-modified DNA was purified with the Wizard DNA Clean-up System and vacuum manifold (Promega, Madison, WI, USA) according to the manufacturer’s manual. .. Freshly prepared NaOH solution was added to a final concentration of 0.3 M, and samples were incubated at 37°C for 15 min, followed by neutralization with ammonium acetate (pH 7.0; final concentration, 3.0 M) and ethanol precipitation.

Article Title: PTEN promoter methylation and activation of the PI3K/Akt/mTOR pathway in pediatric gliomas and influence on clinical outcome
Article Snippet: Bisulfite-modified DNA was purified with the Wizard DNA Clean-up System and vacuum manifold (Promega) according to the manufacturer's manual. .. Freshly prepared NaOH solution was added to a final concentration of 0.3 M, and samples were incubated at 37°C for 15 min, followed by neutralization with ammonium acetate (pH 7.0; final concentration 3.0 M) and ethanol precipitation.

TA Cloning:

Article Title: Upregulation of TRAG3 gene in urothelial carcinoma of the bladder
Article Snippet: Modified samples were purified using Wizard DNA Clean-Up System desalting columns (Promega) followed by ethanol precipitation. .. The PCR products were cloned into TA cloning vector pCR 2.1-TOPO (Invitrogen).

Quantitative RT-PCR:

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: Paragraph title: TaqMan real-time RT-PCR for quantifying avian HEV RNA. ... The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol.

Real-time Polymerase Chain Reaction:

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: For determining the amount of CaHEV, TaqMan real-time RT-PCR was developed and performed with a 20-μl reaction mixture on the StepOnePlus real-time PCR system (Applied Biosystems, Foster City, CA, USA) using the QuantiTect probe RT-PCR kit (Qiagen, Hilden, Germany). .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol.

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: For determining the amount of CaHEV, TaqMan real-time RT-PCR was developed and performed with a 20-μl reaction mixture on the StepOnePlus real-time PCR system (Applied Biosystems, Foster City, CA, USA) using the QuantiTect probe RT-PCR kit (Qiagen, Hilden, Germany). .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol.

Article Title: A TGF-β type II receptor that associates with developmental transition in Haemonchus contortus in vitro
Article Snippet: Preparation of nucleic acids Using a Wizard DNA Clean-Up System (Promega Corporation, USA), genomic DNA was isolated from single female or male adults of H . contortus . .. RNA yields were verified by spectrophotometric analysis (NanoDrop Technologies), cDNA was reverse transcribed from total RNA (1 μg) using the RevertAid First-Strand cDNA Synthesis Kit (Thermo Scientific, USA) to amplify coding sequence, or employing the PrimeScript RT reagent Kit with gDNA Eraser (Perfect Real Time) (Takara, Japan) for real-time PCR.

Incubation:

Article Title: Activation of PI3K/mTOR pathway occurs in most adult low-grade gliomas and predicts patient survival
Article Snippet: Samples were incubated at 55°C for 16 h. Two drops of mineral oil were layered on top of the solution to prevent evaporation. .. Bisulfite-modified DNA was purified with the Wizard DNA Clean-up System and vacuum manifold (Promega, Madison, WI, USA) according to the manufacturer’s manual.

Article Title: PTEN promoter methylation and activation of the PI3K/Akt/mTOR pathway in pediatric gliomas and influence on clinical outcome
Article Snippet: Samples were incubated at 55°C for 16 h. Two drops of mineral oil were layered on top of the solution to prevent evaporation. .. Bisulfite-modified DNA was purified with the Wizard DNA Clean-up System and vacuum manifold (Promega) according to the manufacturer's manual.

Article Title: 2'-Deoxy-N4-[2-(4-nitrophenyl) ethoxycarbonyl]-5-azacytidine: a novel inhibitor of DNA methyltransferase that requires activation by human carboxylesterase 1
Article Snippet: .. The samples were incubated at 50°C for 16 h. The bisulfite-treated DNA was isolated using Wizard DNA Clean-Up System (Promega). ..

Modification:

Article Title: Strategy for the analysis of tissue-specific methylation changes without physical isolation
Article Snippet: Frommer’s protocol [ ] was followed with some modification described in Gonzalez et Al. [ ]. .. Finally, DNA was purified using Wizard DNA Clean Up System (Promega, cat Nº A7280) and then desulfonated by adding 0.3 M NaOH and incubating at 37°C for 20 min. As a control of unmethylated cytosine conversion, we examined a segment of Atp1, a mitochondrial gene that lacks methylation, so that all cytosines are expected to be converted to thymines.

Article Title: Upregulation of TRAG3 gene in urothelial carcinoma of the bladder
Article Snippet: .. Modified samples were purified using Wizard DNA Clean-Up System desalting columns (Promega) followed by ethanol precipitation. .. Bisulfite-modified DNA (100 ng) was PCR amplified (35 cycles: 95 °C, 30 sec; 54 °C, 30 sec; 72 °C, 1 min) with primer sets described previously in a 25-µl reaction mixture.

Article Title: Activation of PI3K/mTOR pathway occurs in most adult low-grade gliomas and predicts patient survival
Article Snippet: Bisulfite modification of genomic DNA was performed as described previously. .. Bisulfite-modified DNA was purified with the Wizard DNA Clean-up System and vacuum manifold (Promega, Madison, WI, USA) according to the manufacturer’s manual.

Article Title: Complete Genome Sequence of Stenotrophomonas Phage Mendera
Article Snippet: .. Phage genomic DNA was purified with the Promega Wizard DNA clean-up system modified according to the shotgun library preparation protocol ( ). .. Libraries were prepared with the Illumina TruSeq Nano low-throughput kit and sequenced via Illumina MiSeq with paired-end 250-bp reads using V2 500-cycle chemistry.

Article Title: Applying DNA barcoding to red macroalgae: a preliminary appraisal holds promise for future applications
Article Snippet: .. DNA was extracted with a protocol modified from (instead of the final agarose gel cleaning procedure, the DNA was purified with the Wizard® DNA Clean-Up System, Promega Corp., Madison, WI). .. The cox1 sequences for Cyanidium caldarium (Tilden) Geitler , Cyanidioschyzon merolae P. De Luca, R. Taddei et L. Varano (NC 000887), Chondrus crispus Stackhouse (NC 001677), and Porphyra purpurea (Roth) C. Agardh (NC 002007) were acquired from GenBank and aligned by eye in MacClade 4 (v. 4.06) for OSX ( ).

Article Title: Stool-based DNA testing, a new noninvasive method for colorectal cancer screening, the first report from Iran
Article Snippet: .. Modified DNA was purified using a Promega Wizard DNA Clean-Up System, according to the manufacturer’s instructions and then was stored at -20°C until it was used for PCR. .. Modified DNA was amplified using primers specific for methylated and unmethylated p 16 sequences as previously described[ ].

Article Title: PTEN promoter methylation and activation of the PI3K/Akt/mTOR pathway in pediatric gliomas and influence on clinical outcome
Article Snippet: Bisulfite-modified DNA was purified with the Wizard DNA Clean-up System and vacuum manifold (Promega) according to the manufacturer's manual. .. Normal human peripheral blood lymphocyte DNA samples, treated and untreated with DNA methylase (M. Sss I; New England BioLabs), were also modified as positive and negative controls, respectively.

Article Title: 2'-Deoxy-N4-[2-(4-nitrophenyl) ethoxycarbonyl]-5-azacytidine: a novel inhibitor of DNA methyltransferase that requires activation by human carboxylesterase 1
Article Snippet: Bisulfite modification of genomic DNA has been described previously [ ]. .. The samples were incubated at 50°C for 16 h. The bisulfite-treated DNA was isolated using Wizard DNA Clean-Up System (Promega).

Electron Microscopy:

Article Title: Complete Genome Sequence of Stenotrophomonas Phage Mendera
Article Snippet: Mendera myophage morphology was identified via negative-stain transmission electron microscopy with 2% (wt/vol) uranyl acetate at the Texas A & M Microscopy and Imaging Center ( ). .. Phage genomic DNA was purified with the Promega Wizard DNA clean-up system modified according to the shotgun library preparation protocol ( ).

Sequencing:

Article Title: Complete Genome Sequence of Stenotrophomonas Phage Mendera
Article Snippet: Phage genomic DNA was purified with the Promega Wizard DNA clean-up system modified according to the shotgun library preparation protocol ( ). .. The 222,492 total sequence reads were quality controlled with FastQC ( www.bioinformatics.babraham.ac.uk/projects/fastqc ), trimmed using the FastX toolkit v0.0.14 ( http://hannonlab.cshl.edu/fastx_toolkit/ ), and assembled into a single raw contig at 11.6-fold coverage using SPAdes v3.5.0 at default parameters ( ).

Article Title: A TGF-β type II receptor that associates with developmental transition in Haemonchus contortus in vitro
Article Snippet: Preparation of nucleic acids Using a Wizard DNA Clean-Up System (Promega Corporation, USA), genomic DNA was isolated from single female or male adults of H . contortus . .. RNA yields were verified by spectrophotometric analysis (NanoDrop Technologies), cDNA was reverse transcribed from total RNA (1 μg) using the RevertAid First-Strand cDNA Synthesis Kit (Thermo Scientific, USA) to amplify coding sequence, or employing the PrimeScript RT reagent Kit with gDNA Eraser (Perfect Real Time) (Takara, Japan) for real-time PCR.

Concentration Assay:

Article Title: Upregulation of TRAG3 gene in urothelial carcinoma of the bladder
Article Snippet: Briefly, 1–2 µg (5–10 µl) genomic DNA was denatured by NaOH (final concentration, 0.2 M), 30 µl of 10 mM hydroquinone (Sigma), and 520 µl of 3M sodium bisulfite (Sigma) at pH 5 and 50 °C for 8 hrs. .. Modified samples were purified using Wizard DNA Clean-Up System desalting columns (Promega) followed by ethanol precipitation.

Article Title: Activation of PI3K/mTOR pathway occurs in most adult low-grade gliomas and predicts patient survival
Article Snippet: Bisulfite-modified DNA was purified with the Wizard DNA Clean-up System and vacuum manifold (Promega, Madison, WI, USA) according to the manufacturer’s manual. .. Freshly prepared NaOH solution was added to a final concentration of 0.3 M, and samples were incubated at 37°C for 15 min, followed by neutralization with ammonium acetate (pH 7.0; final concentration, 3.0 M) and ethanol precipitation.

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol. .. The concentration of RNA was determined at least four times by using a spectrophotometer (BioTek Instruments, Inc., Belgium, WI, USA), and the numbers of copies per microliter were calculated using the mean values and the formula [(grams/microliter RNA)/(length × 340)] × 6.022 × 1023 , where length is the number of nucleotides.

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol. .. The concentration of RNA was determined at least four times by using a spectrophotometer (BioTek Instruments, Inc., Belgium, WI, USA), and the numbers of copies per microliter were calculated using the mean values and the formula [(grams/microliter RNA)/(length × 340)] × 6.022 × 1023 , where length is the number of nucleotides.

Article Title: PTEN promoter methylation and activation of the PI3K/Akt/mTOR pathway in pediatric gliomas and influence on clinical outcome
Article Snippet: Bisulfite-modified DNA was purified with the Wizard DNA Clean-up System and vacuum manifold (Promega) according to the manufacturer's manual. .. Freshly prepared NaOH solution was added to a final concentration of 0.3 M, and samples were incubated at 37°C for 15 min, followed by neutralization with ammonium acetate (pH 7.0; final concentration 3.0 M) and ethanol precipitation.

Genomic Sequencing:

Article Title: Upregulation of TRAG3 gene in urothelial carcinoma of the bladder
Article Snippet: Paragraph title: Bisulfite Genomic Sequencing and Methylation specific PCR (MSP) Assay ... Modified samples were purified using Wizard DNA Clean-Up System desalting columns (Promega) followed by ethanol precipitation.

Infection:

Article Title: Complete Genome Sequence of Stenotrophomonas Phage Mendera
Article Snippet: As S. maltophilia is an opportunist, infection caused by the bacterium is a major problem for cystic fibrosis patients ( ). .. Phage genomic DNA was purified with the Promega Wizard DNA clean-up system modified according to the shotgun library preparation protocol ( ).

Imaging:

Article Title: Complete Genome Sequence of Stenotrophomonas Phage Mendera
Article Snippet: Mendera myophage morphology was identified via negative-stain transmission electron microscopy with 2% (wt/vol) uranyl acetate at the Texas A & M Microscopy and Imaging Center ( ). .. Phage genomic DNA was purified with the Promega Wizard DNA clean-up system modified according to the shotgun library preparation protocol ( ).

Transmission Assay:

Article Title: Complete Genome Sequence of Stenotrophomonas Phage Mendera
Article Snippet: Mendera myophage morphology was identified via negative-stain transmission electron microscopy with 2% (wt/vol) uranyl acetate at the Texas A & M Microscopy and Imaging Center ( ). .. Phage genomic DNA was purified with the Promega Wizard DNA clean-up system modified according to the shotgun library preparation protocol ( ).

Polymerase Chain Reaction:

Article Title: Upregulation of TRAG3 gene in urothelial carcinoma of the bladder
Article Snippet: Paragraph title: Bisulfite Genomic Sequencing and Methylation specific PCR (MSP) Assay ... Modified samples were purified using Wizard DNA Clean-Up System desalting columns (Promega) followed by ethanol precipitation.

Article Title: Complete Genome Sequence of Stenotrophomonas Phage Mendera
Article Snippet: Phage genomic DNA was purified with the Promega Wizard DNA clean-up system modified according to the shotgun library preparation protocol ( ). .. Contig accuracy and completeness were confirmed by PCR (forward primer, 5′-GTCACCGCGACTACGATAAG-3′; reverse primer, 5′-GAACGACAGCGAGCATACA-3′) off the contig ends and Sanger sequencing of the product.

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: The cDNA from RNA was synthesized via reverse transcription for 30 min at 50°C, followed by an PCR initial activation step for 15 min at 95°C. .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol.

Article Title: Applying DNA barcoding to red macroalgae: a preliminary appraisal holds promise for future applications
Article Snippet: DNA was extracted with a protocol modified from (instead of the final agarose gel cleaning procedure, the DNA was purified with the Wizard® DNA Clean-Up System, Promega Corp., Madison, WI). .. The PCR amplification profile followed , but using an annealing temperature of 50 °C.

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: The cDNA from RNA was synthesized via reverse transcription for 30 min at 50°C, followed by an PCR initial activation step for 15 min at 95°C. .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol.

Article Title: Stool-based DNA testing, a new noninvasive method for colorectal cancer screening, the first report from Iran
Article Snippet: .. Modified DNA was purified using a Promega Wizard DNA Clean-Up System, according to the manufacturer’s instructions and then was stored at -20°C until it was used for PCR. .. Modified DNA was amplified using primers specific for methylated and unmethylated p 16 sequences as previously described[ ].

Molecular Weight:

Article Title: Upregulation of TRAG3 gene in urothelial carcinoma of the bladder
Article Snippet: High molecular weight genomic DNA was obtained from the indicated cell lines and subjected to bisulfite modification as previously described . .. Modified samples were purified using Wizard DNA Clean-Up System desalting columns (Promega) followed by ethanol precipitation.

DNA Extraction:

Article Title: High Bacterial Diversity in Permanently Cold Marine Sediments
Article Snippet: Paragraph title: DNA extraction and purification. ... The crude DNA was purified with the WIZARD DNA Clean Up System (Promega, Madison, Wis.).

Article Title: PTEN promoter methylation and activation of the PI3K/Akt/mTOR pathway in pediatric gliomas and influence on clinical outcome
Article Snippet: DNA isolation and bisulfite treatment were carried out for samples for which frozen tissues were available, as previously described. .. Bisulfite-modified DNA was purified with the Wizard DNA Clean-up System and vacuum manifold (Promega) according to the manufacturer's manual.

Methylation:

Article Title: Strategy for the analysis of tissue-specific methylation changes without physical isolation
Article Snippet: .. Finally, DNA was purified using Wizard DNA Clean Up System (Promega, cat Nº A7280) and then desulfonated by adding 0.3 M NaOH and incubating at 37°C for 20 min. As a control of unmethylated cytosine conversion, we examined a segment of Atp1, a mitochondrial gene that lacks methylation, so that all cytosines are expected to be converted to thymines. ..

Article Title: Upregulation of TRAG3 gene in urothelial carcinoma of the bladder
Article Snippet: Paragraph title: Bisulfite Genomic Sequencing and Methylation specific PCR (MSP) Assay ... Modified samples were purified using Wizard DNA Clean-Up System desalting columns (Promega) followed by ethanol precipitation.

Article Title: Stool-based DNA testing, a new noninvasive method for colorectal cancer screening, the first report from Iran
Article Snippet: Paragraph title: p16 methylation analysis ... Modified DNA was purified using a Promega Wizard DNA Clean-Up System, according to the manufacturer’s instructions and then was stored at -20°C until it was used for PCR.

Isolation:

Article Title: Complete Genome Sequence of Stenotrophomonas Phage Mendera
Article Snippet: For isolation, the host S. maltophilia (ATCC 17807) was grown aerobically at 30°C in nutrient broth or agar (BD), and phage was propagated by the soft-agar overlay method ( ). .. Phage genomic DNA was purified with the Promega Wizard DNA clean-up system modified according to the shotgun library preparation protocol ( ).

Article Title: A TGF-β type II receptor that associates with developmental transition in Haemonchus contortus in vitro
Article Snippet: .. Preparation of nucleic acids Using a Wizard DNA Clean-Up System (Promega Corporation, USA), genomic DNA was isolated from single female or male adults of H . contortus . .. Using the TRIzol reagent extraction method (Life Technologies, USA), total RNA was isolated from pools of 100–500 individuals of each developmental stage (i.e. egg, L1, L2, L3, female L4, male L4, female adult and male adult stages) of H . contortus .

Article Title: PTEN promoter methylation and activation of the PI3K/Akt/mTOR pathway in pediatric gliomas and influence on clinical outcome
Article Snippet: Briefly, genomic DNA was isolated from approximately 25 mg wet weight of frozen tissue using the QIAamp DNA Mini Kit (Qiagen) according to the manufacturer's instructions. .. Bisulfite-modified DNA was purified with the Wizard DNA Clean-up System and vacuum manifold (Promega) according to the manufacturer's manual.

Article Title: 2'-Deoxy-N4-[2-(4-nitrophenyl) ethoxycarbonyl]-5-azacytidine: a novel inhibitor of DNA methyltransferase that requires activation by human carboxylesterase 1
Article Snippet: .. The samples were incubated at 50°C for 16 h. The bisulfite-treated DNA was isolated using Wizard DNA Clean-Up System (Promega). ..

Size-exclusion Chromatography:

Article Title: Upregulation of TRAG3 gene in urothelial carcinoma of the bladder
Article Snippet: Modified samples were purified using Wizard DNA Clean-Up System desalting columns (Promega) followed by ethanol precipitation. .. Bisulfite-modified DNA (100 ng) was PCR amplified (35 cycles: 95 °C, 30 sec; 54 °C, 30 sec; 72 °C, 1 min) with primer sets described previously in a 25-µl reaction mixture.

Microscopy:

Article Title: Complete Genome Sequence of Stenotrophomonas Phage Mendera
Article Snippet: Mendera myophage morphology was identified via negative-stain transmission electron microscopy with 2% (wt/vol) uranyl acetate at the Texas A & M Microscopy and Imaging Center ( ). .. Phage genomic DNA was purified with the Promega Wizard DNA clean-up system modified according to the shotgun library preparation protocol ( ).

Purification:

Article Title: Strategy for the analysis of tissue-specific methylation changes without physical isolation
Article Snippet: .. Finally, DNA was purified using Wizard DNA Clean Up System (Promega, cat Nº A7280) and then desulfonated by adding 0.3 M NaOH and incubating at 37°C for 20 min. As a control of unmethylated cytosine conversion, we examined a segment of Atp1, a mitochondrial gene that lacks methylation, so that all cytosines are expected to be converted to thymines. ..

Article Title: Upregulation of TRAG3 gene in urothelial carcinoma of the bladder
Article Snippet: .. Modified samples were purified using Wizard DNA Clean-Up System desalting columns (Promega) followed by ethanol precipitation. .. Bisulfite-modified DNA (100 ng) was PCR amplified (35 cycles: 95 °C, 30 sec; 54 °C, 30 sec; 72 °C, 1 min) with primer sets described previously in a 25-µl reaction mixture.

Article Title: Activation of PI3K/mTOR pathway occurs in most adult low-grade gliomas and predicts patient survival
Article Snippet: .. Bisulfite-modified DNA was purified with the Wizard DNA Clean-up System and vacuum manifold (Promega, Madison, WI, USA) according to the manufacturer’s manual. .. Freshly prepared NaOH solution was added to a final concentration of 0.3 M, and samples were incubated at 37°C for 15 min, followed by neutralization with ammonium acetate (pH 7.0; final concentration, 3.0 M) and ethanol precipitation.

Article Title: Complete Genome Sequence of Stenotrophomonas Phage Mendera
Article Snippet: .. Phage genomic DNA was purified with the Promega Wizard DNA clean-up system modified according to the shotgun library preparation protocol ( ). .. Libraries were prepared with the Illumina TruSeq Nano low-throughput kit and sequenced via Illumina MiSeq with paired-end 250-bp reads using V2 500-cycle chemistry.

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol. .. Next, the purified plasmid DNA was transcribed in vitro using a RiboMAX T7 large-scale RNA production system (Promega, Madison, WI, USA).

Article Title: Applying DNA barcoding to red macroalgae: a preliminary appraisal holds promise for future applications
Article Snippet: .. DNA was extracted with a protocol modified from (instead of the final agarose gel cleaning procedure, the DNA was purified with the Wizard® DNA Clean-Up System, Promega Corp., Madison, WI). .. The cox1 sequences for Cyanidium caldarium (Tilden) Geitler , Cyanidioschyzon merolae P. De Luca, R. Taddei et L. Varano (NC 000887), Chondrus crispus Stackhouse (NC 001677), and Porphyra purpurea (Roth) C. Agardh (NC 002007) were acquired from GenBank and aligned by eye in MacClade 4 (v. 4.06) for OSX ( ).

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol. .. Next, the purified plasmid DNA was transcribed in vitro using a RiboMAX T7 large-scale RNA production system (Promega, Madison, WI, USA).

Article Title: High Bacterial Diversity in Permanently Cold Marine Sediments
Article Snippet: .. The crude DNA was purified with the WIZARD DNA Clean Up System (Promega, Madison, Wis.). ..

Article Title: Stool-based DNA testing, a new noninvasive method for colorectal cancer screening, the first report from Iran
Article Snippet: .. Modified DNA was purified using a Promega Wizard DNA Clean-Up System, according to the manufacturer’s instructions and then was stored at -20°C until it was used for PCR. .. Modified DNA was amplified using primers specific for methylated and unmethylated p 16 sequences as previously described[ ].

Article Title: PTEN promoter methylation and activation of the PI3K/Akt/mTOR pathway in pediatric gliomas and influence on clinical outcome
Article Snippet: .. Bisulfite-modified DNA was purified with the Wizard DNA Clean-up System and vacuum manifold (Promega) according to the manufacturer's manual. .. Freshly prepared NaOH solution was added to a final concentration of 0.3 M, and samples were incubated at 37°C for 15 min, followed by neutralization with ammonium acetate (pH 7.0; final concentration 3.0 M) and ethanol precipitation.

Reverse Transcription Polymerase Chain Reaction:

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: Next, targeting ORF3 was amplified for 45 cycles at 94°C for 15 s and 60°C for 60 s. In addition, to generate the internal control RNA, a 174-bp RT-PCR product was amplified using the primers Taqman-T7f and Taqman-R ( ) and then cloned into the pMD19-T vector (TaKaRa Biotech Corporation, Dalian, China). .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol.

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: In addition, to generate the internal control RNA, a 174-bp RT-PCR product was amplified using the primers Taqman-T7f and Taqman-R ( ) and then cloned into the pMD19-T vector (TaKaRa Biotech Corporation, Dalian, China). .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol.

MSP Assay:

Article Title: Upregulation of TRAG3 gene in urothelial carcinoma of the bladder
Article Snippet: Paragraph title: Bisulfite Genomic Sequencing and Methylation specific PCR (MSP) Assay ... Modified samples were purified using Wizard DNA Clean-Up System desalting columns (Promega) followed by ethanol precipitation.

Plasmid Preparation:

Article Title: Upregulation of TRAG3 gene in urothelial carcinoma of the bladder
Article Snippet: Modified samples were purified using Wizard DNA Clean-Up System desalting columns (Promega) followed by ethanol precipitation. .. The PCR products were cloned into TA cloning vector pCR 2.1-TOPO (Invitrogen).

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: Next, targeting ORF3 was amplified for 45 cycles at 94°C for 15 s and 60°C for 60 s. In addition, to generate the internal control RNA, a 174-bp RT-PCR product was amplified using the primers Taqman-T7f and Taqman-R ( ) and then cloned into the pMD19-T vector (TaKaRa Biotech Corporation, Dalian, China). .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol.

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: In addition, to generate the internal control RNA, a 174-bp RT-PCR product was amplified using the primers Taqman-T7f and Taqman-R ( ) and then cloned into the pMD19-T vector (TaKaRa Biotech Corporation, Dalian, China). .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol.

Agarose Gel Electrophoresis:

Article Title: Applying DNA barcoding to red macroalgae: a preliminary appraisal holds promise for future applications
Article Snippet: .. DNA was extracted with a protocol modified from (instead of the final agarose gel cleaning procedure, the DNA was purified with the Wizard® DNA Clean-Up System, Promega Corp., Madison, WI). .. The cox1 sequences for Cyanidium caldarium (Tilden) Geitler , Cyanidioschyzon merolae P. De Luca, R. Taddei et L. Varano (NC 000887), Chondrus crispus Stackhouse (NC 001677), and Porphyra purpurea (Roth) C. Agardh (NC 002007) were acquired from GenBank and aligned by eye in MacClade 4 (v. 4.06) for OSX ( ).

Article Title: High Bacterial Diversity in Permanently Cold Marine Sediments
Article Snippet: The crude DNA was purified with the WIZARD DNA Clean Up System (Promega, Madison, Wis.). .. High-molecular-weight DNA was cut out of an agarose gel and extracted with a GeneClean II Kit (Bio 101 Inc., La Jolla, Calif.) by following the manufacturer’s instructions.

In Vitro:

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol. .. Next, the purified plasmid DNA was transcribed in vitro using a RiboMAX T7 large-scale RNA production system (Promega, Madison, WI, USA).

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol. .. Next, the purified plasmid DNA was transcribed in vitro using a RiboMAX T7 large-scale RNA production system (Promega, Madison, WI, USA).

Ethanol Precipitation:

Article Title: Upregulation of TRAG3 gene in urothelial carcinoma of the bladder
Article Snippet: .. Modified samples were purified using Wizard DNA Clean-Up System desalting columns (Promega) followed by ethanol precipitation. .. Bisulfite-modified DNA (100 ng) was PCR amplified (35 cycles: 95 °C, 30 sec; 54 °C, 30 sec; 72 °C, 1 min) with primer sets described previously in a 25-µl reaction mixture.

Article Title: Activation of PI3K/mTOR pathway occurs in most adult low-grade gliomas and predicts patient survival
Article Snippet: Bisulfite-modified DNA was purified with the Wizard DNA Clean-up System and vacuum manifold (Promega, Madison, WI, USA) according to the manufacturer’s manual. .. Freshly prepared NaOH solution was added to a final concentration of 0.3 M, and samples were incubated at 37°C for 15 min, followed by neutralization with ammonium acetate (pH 7.0; final concentration, 3.0 M) and ethanol precipitation.

Article Title: PTEN promoter methylation and activation of the PI3K/Akt/mTOR pathway in pediatric gliomas and influence on clinical outcome
Article Snippet: Bisulfite-modified DNA was purified with the Wizard DNA Clean-up System and vacuum manifold (Promega) according to the manufacturer's manual. .. Freshly prepared NaOH solution was added to a final concentration of 0.3 M, and samples were incubated at 37°C for 15 min, followed by neutralization with ammonium acetate (pH 7.0; final concentration 3.0 M) and ethanol precipitation.

Spectrophotometry:

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol. .. The concentration of RNA was determined at least four times by using a spectrophotometer (BioTek Instruments, Inc., Belgium, WI, USA), and the numbers of copies per microliter were calculated using the mean values and the formula [(grams/microliter RNA)/(length × 340)] × 6.022 × 1023 , where length is the number of nucleotides.

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol. .. The concentration of RNA was determined at least four times by using a spectrophotometer (BioTek Instruments, Inc., Belgium, WI, USA), and the numbers of copies per microliter were calculated using the mean values and the formula [(grams/microliter RNA)/(length × 340)] × 6.022 × 1023 , where length is the number of nucleotides.

Evaporation:

Article Title: Activation of PI3K/mTOR pathway occurs in most adult low-grade gliomas and predicts patient survival
Article Snippet: Samples were incubated at 55°C for 16 h. Two drops of mineral oil were layered on top of the solution to prevent evaporation. .. Bisulfite-modified DNA was purified with the Wizard DNA Clean-up System and vacuum manifold (Promega, Madison, WI, USA) according to the manufacturer’s manual.

Article Title: PTEN promoter methylation and activation of the PI3K/Akt/mTOR pathway in pediatric gliomas and influence on clinical outcome
Article Snippet: Samples were incubated at 55°C for 16 h. Two drops of mineral oil were layered on top of the solution to prevent evaporation. .. Bisulfite-modified DNA was purified with the Wizard DNA Clean-up System and vacuum manifold (Promega) according to the manufacturer's manual.

Activation Assay:

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: The cDNA from RNA was synthesized via reverse transcription for 30 min at 50°C, followed by an PCR initial activation step for 15 min at 95°C. .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol.

Article Title: Chicken Organic Anion-Transporting Polypeptide 1A2, a Novel Avian Hepatitis E Virus (HEV) ORF2-Interacting Protein, Is Involved in Avian HEV Infection
Article Snippet: The cDNA from RNA was synthesized via reverse transcription for 30 min at 50°C, followed by an PCR initial activation step for 15 min at 95°C. .. The vectors were linearized using the PstI restriction enzyme (TaKaRa Biotech Corporation, Dalian, China) and purified by using the Wizard DNA clean-up system (Promega, Madison, WI, USA), according to the manufacturer’s protocol.

Lysis:

Article Title: High Bacterial Diversity in Permanently Cold Marine Sediments
Article Snippet: The protocol encompassed three cycles of freezing and thawing, chemical lysis in a high-salt extraction buffer (1.5 M NaCl) by heating of the suspension in the presence of sodium dodecyl sulfate (SDS) and hexadecyltrimethylammonium bromide, and a proteinase K step. .. The crude DNA was purified with the WIZARD DNA Clean Up System (Promega, Madison, Wis.).

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  • 99
    Promega dna wizard clean up system
    Results of the comparison of the two best methods on microarray . The mean value and standard error of four individual spots on a microarray are expressed in arbitrary fluorescence units on the y-axis. On the x-axis the number of the experiment, the number of the <t>DNA</t> mix and the weight percentage of the genomic DNA is given for A: bar , B: cry1Ab , C: hmg and D: <t>TC1507</t> detection. * indicates a p-value of
    Dna Wizard Clean Up System, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna wizard clean up system/product/Promega
    Average 99 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    dna wizard clean up system - by Bioz Stars, 2020-04
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    99
    Promega wizard pcr preps dna purification system
    Restriction endonuclease digestion patterns for the A3′ amplicon of the CROP region of the toxin A gene for HSC98. The <t>PCR</t> amplicon product A3′ (lane 2) obtained from HSC98 using the A3 primer set (product was 1,250 bp) was exposed to Spe I (lane 3) and Eco I (lane 4) restriction endonucleases to determine restriction fragment length patterns. The A3′ amplicon (lane 2) had no restriction site for Eco RI (lane 4) and had one restriction site for Spe I (lane 3). A 500-bp <t>DNA</t> ladder is shown in lanes 1 and 6, and a 100-bp ladder is shown in lane 5.
    Wizard Pcr Preps Dna Purification System, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 204 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/wizard pcr preps dna purification system/product/Promega
    Average 99 stars, based on 204 article reviews
    Price from $9.99 to $1999.99
    wizard pcr preps dna purification system - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

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    Results of the comparison of the two best methods on microarray . The mean value and standard error of four individual spots on a microarray are expressed in arbitrary fluorescence units on the y-axis. On the x-axis the number of the experiment, the number of the DNA mix and the weight percentage of the genomic DNA is given for A: bar , B: cry1Ab , C: hmg and D: TC1507 detection. * indicates a p-value of

    Journal: BMC Biotechnology

    Article Title: Comparison and transfer testing of multiplex ligation detection methods for GM plants

    doi: 10.1186/1472-6750-12-4

    Figure Lengend Snippet: Results of the comparison of the two best methods on microarray . The mean value and standard error of four individual spots on a microarray are expressed in arbitrary fluorescence units on the y-axis. On the x-axis the number of the experiment, the number of the DNA mix and the weight percentage of the genomic DNA is given for A: bar , B: cry1Ab , C: hmg and D: TC1507 detection. * indicates a p-value of

    Article Snippet: For GTS 40-3-2, 100% Bt176 and 100% TC1507 the DNA Wizard Clean up system for genomic DNA (Promega) was used.

    Techniques: Microarray, Fluorescence

    Restriction endonuclease digestion patterns for the A3′ amplicon of the CROP region of the toxin A gene for HSC98. The PCR amplicon product A3′ (lane 2) obtained from HSC98 using the A3 primer set (product was 1,250 bp) was exposed to Spe I (lane 3) and Eco I (lane 4) restriction endonucleases to determine restriction fragment length patterns. The A3′ amplicon (lane 2) had no restriction site for Eco RI (lane 4) and had one restriction site for Spe I (lane 3). A 500-bp DNA ladder is shown in lanes 1 and 6, and a 100-bp ladder is shown in lane 5.

    Journal: Journal of Clinical Microbiology

    Article Title: Characterization of a Toxin A-Negative, Toxin B-Positive Strain of Clostridium difficile Responsible for a Nosocomial Outbreak of Clostridium difficile-Associated Diarrhea

    doi:

    Figure Lengend Snippet: Restriction endonuclease digestion patterns for the A3′ amplicon of the CROP region of the toxin A gene for HSC98. The PCR amplicon product A3′ (lane 2) obtained from HSC98 using the A3 primer set (product was 1,250 bp) was exposed to Spe I (lane 3) and Eco I (lane 4) restriction endonucleases to determine restriction fragment length patterns. The A3′ amplicon (lane 2) had no restriction site for Eco RI (lane 4) and had one restriction site for Spe I (lane 3). A 500-bp DNA ladder is shown in lanes 1 and 6, and a 100-bp ladder is shown in lane 5.

    Article Snippet: The amplified product from this PCR protocol was purified using the Wizard PCR Preps DNA purification system (Promega Corp., Madison, Wis.), and this purified preparation was used for DNA sequencing.

    Techniques: Amplification, Polymerase Chain Reaction

    UP-PCR results of some patients. We have detected all the patients and finally we chose some positive bacteria and carried out the electrophoresis, so every line had PCR products. The deeper the stripe, the more the bacteria. Lines 1–9 represented PCR product of the 16S rRNA gene of Staphylococcus aureus , Staphylococcus epidermidis , Pseudomonas aeruginosa , Escherichia coli , Streptococcus viridans , Staphylococcus saprophyticus, Corynebacterium parvum , Klebsiella pneumoniae , and Enterobacter cloacae ; M for DL2000 DNA Marker; ten for negative control.

    Journal: BioMed Research International

    Article Title: Genetic Identification and Risk Factor Analysis of Asymptomatic Bacterial Colonization on Cardiovascular Implantable Electronic Devices

    doi: 10.1155/2014/725163

    Figure Lengend Snippet: UP-PCR results of some patients. We have detected all the patients and finally we chose some positive bacteria and carried out the electrophoresis, so every line had PCR products. The deeper the stripe, the more the bacteria. Lines 1–9 represented PCR product of the 16S rRNA gene of Staphylococcus aureus , Staphylococcus epidermidis , Pseudomonas aeruginosa , Escherichia coli , Streptococcus viridans , Staphylococcus saprophyticus, Corynebacterium parvum , Klebsiella pneumoniae , and Enterobacter cloacae ; M for DL2000 DNA Marker; ten for negative control.

    Article Snippet: The PCR product was purified using Wizard PCR Preps DNA Purification System (Promega) and then ligated into the pGEM-T Easy Vector (Promega).

    Techniques: Polymerase Chain Reaction, Electrophoresis, Marker, Negative Control

    The MALS mutagenesis procedure . Target DNA is individually amplified with two pairs of oligonucleotides SO1/IR1 and SO2/IF1. IF1 and IR1 are phosphorylated (rose circles), while SO1 and SO2 are dephosphorylated. The desired mutation is incorporated at the 5' end of IR1 oligonucleotide (shown in red). Panel 1 on the right shows the types of mutations available with MALS. PCR-generated DNA fragments are ligated. The resulting DNA population consists of homomeric ligation products (type A), non-ligated DNA fragments (type B), and molecules representing full length target DNA (type C) (panel 2 on the right). The entire DNA population is then amplified with suppression oligonucleotides SO1 and SO2. Intramolecular hybridization of inverted repeat sequences prevents efficient replication of type A molecules, while type B molecules amplify linearly (panel 3 on the right). Only heteromeric ligation products (type C) amplify exponentially. The resulting DNA population predominantly consists of type C molecules. An aliquot of the PCR mixture is used for the next round of mutagenesis employing internal oligonucleotides specific for another region of target DNA. Finally, SO1 and SO2 suppression sequences (green and blue, respectively) are digested with restriction endonucleases and DNA fragments are ligated with linearized vector for subsequent subcloning.

    Journal: BMC Biotechnology

    Article Title: MALS: an efficient strategy for multiple site-directed mutagenesis employing a combination of DNA amplification, ligation and suppression PCR

    doi: 10.1186/1472-6750-9-83

    Figure Lengend Snippet: The MALS mutagenesis procedure . Target DNA is individually amplified with two pairs of oligonucleotides SO1/IR1 and SO2/IF1. IF1 and IR1 are phosphorylated (rose circles), while SO1 and SO2 are dephosphorylated. The desired mutation is incorporated at the 5' end of IR1 oligonucleotide (shown in red). Panel 1 on the right shows the types of mutations available with MALS. PCR-generated DNA fragments are ligated. The resulting DNA population consists of homomeric ligation products (type A), non-ligated DNA fragments (type B), and molecules representing full length target DNA (type C) (panel 2 on the right). The entire DNA population is then amplified with suppression oligonucleotides SO1 and SO2. Intramolecular hybridization of inverted repeat sequences prevents efficient replication of type A molecules, while type B molecules amplify linearly (panel 3 on the right). Only heteromeric ligation products (type C) amplify exponentially. The resulting DNA population predominantly consists of type C molecules. An aliquot of the PCR mixture is used for the next round of mutagenesis employing internal oligonucleotides specific for another region of target DNA. Finally, SO1 and SO2 suppression sequences (green and blue, respectively) are digested with restriction endonucleases and DNA fragments are ligated with linearized vector for subsequent subcloning.

    Article Snippet: After amplification, the resulting DNA fragments were extracted from PCR mixtures using Wizard PCR Preps DNA Purification System (Promega, Madison, USA).

    Techniques: Mutagenesis, Amplification, Polymerase Chain Reaction, Generated, Ligation, Hybridization, Plasmid Preparation, Subcloning

    Agarose gel electrophoregram demonstrating the types of molecules forming during one individual round of mutagenesis . Lanes 1 and 2, PCR of target DNA with SO1/IR1 and SO2/IF1 oligonucleotides; Lane 3, homomeric (type A) and heteromeric (type C) DNA molecules formed during ligation of SO1/IR1 and SO2/IF1 DNA fragments (also, see Figure 1); Lane 4, suppression PCR with SO1 and SO2 oligonucleotides. The scheme on the right shows a graphical representation of type A and type C molecules that are present in lane 3 of the gel. During suppression PCR, intramolecular hybridization of inverted repeat sequences prevents binding of SO1 and SO2 oligonucleotides to the type A molecules that prevent their effective replication. Type C molecules amplify exponentially (see Additional File 1 for further details). Lane M, DNA size ladder with indicated positions of 1, 2, 3, 4, 6, and 10 kb DNA fragments (GeneRuler 1 kb DNA Ladder, Fermentas).

    Journal: BMC Biotechnology

    Article Title: MALS: an efficient strategy for multiple site-directed mutagenesis employing a combination of DNA amplification, ligation and suppression PCR

    doi: 10.1186/1472-6750-9-83

    Figure Lengend Snippet: Agarose gel electrophoregram demonstrating the types of molecules forming during one individual round of mutagenesis . Lanes 1 and 2, PCR of target DNA with SO1/IR1 and SO2/IF1 oligonucleotides; Lane 3, homomeric (type A) and heteromeric (type C) DNA molecules formed during ligation of SO1/IR1 and SO2/IF1 DNA fragments (also, see Figure 1); Lane 4, suppression PCR with SO1 and SO2 oligonucleotides. The scheme on the right shows a graphical representation of type A and type C molecules that are present in lane 3 of the gel. During suppression PCR, intramolecular hybridization of inverted repeat sequences prevents binding of SO1 and SO2 oligonucleotides to the type A molecules that prevent their effective replication. Type C molecules amplify exponentially (see Additional File 1 for further details). Lane M, DNA size ladder with indicated positions of 1, 2, 3, 4, 6, and 10 kb DNA fragments (GeneRuler 1 kb DNA Ladder, Fermentas).

    Article Snippet: After amplification, the resulting DNA fragments were extracted from PCR mixtures using Wizard PCR Preps DNA Purification System (Promega, Madison, USA).

    Techniques: Agarose Gel Electrophoresis, Mutagenesis, Polymerase Chain Reaction, Ligation, Hybridization, Binding Assay