taq dna polymerase  (Qiagen)

Journal: Journal of Virology

Article Title: Transgenically Expressed T-Rep of Tomato Yellow Leaf Curl Sardinia Virus Acts as a trans-Dominant-Negative Mutant, Inhibiting Viral Transcription and Replication

doi: 10.1128/JVI.75.22.10573-10581.2001

Figure Lengend Snippet: Slowly migrating DNAs are converted into ocDNA by Taq (A) or T4 (B) DNA polymerase treatment. The blots were hybridized with a C1-sense RNA probe. The positions of ocDNA, linear DNA (linDNA), scDNA, and cssDNA forms of viral DNA are indicated. Slowly migrating viral DNAs are indicated with an asterisk (∗). (A) TNAs were extracted from wt protoplasts at 72 h posttransfection with pTOM6 alone (the two lanes on the right) or together with pTOM100C4(−) or pTOM100NT and analyzed directly (−) or following incubation with Taq DNA polymerase for the time indicated below. (B) TNAs were extracted from wt or transgenic (102.22) protoplasts at 72 h posttransfection with pSP97 (TYLCSV-ES[1]) and analyzed following a 1-h incubation with (+) or without (−) T4 DNA polymerase. Lane C, TNAs from a TYLCSV-infected tomato plant digested with Bgl II to show migration of linear DNA.

Article Snippet: To verify the nature of these slowly migrating molecules, TNAs extracted from the samples at 24 and 72 h were incubated with Taq DNA polymerase and analyzed by Southern blot (Fig. B).

Techniques: Incubation, Transgenic Assay, Infection, Migration

Journal: Journal of Virology

Article Title: Transgenically Expressed T-Rep of Tomato Yellow Leaf Curl Sardinia Virus Acts as a trans-Dominant-Negative Mutant, Inhibiting Viral Transcription and Replication

doi: 10.1128/JVI.75.22.10573-10581.2001

Figure Lengend Snippet: Slowly migrating DNAs also accumulate at early stages after transfection of wt protoplasts with TYLCSV. TNAs were extracted from protoplasts transfected with pTOM6. The blots were hybridized with a C1-sense RNA probe. The positions of ocDNA, linear DNA (linDNA), scDNA, and cssDNA forms of viral DNA are indicated. Slow-migrating viral DNAs are indicated with an asterisk (∗). HPT, hours posttransfection. (A) Time course analysis of viral DNA forms. Sample at 72 h was diluted 20-fold to give signals of satisfactory intensity on the autoradiographic film. (B) Taq DNA polymerase treatment of TNAs at 24 and 72 h after transfection. Samples were incubated for 10 min at 37°C with (+) or without (−) the polymerase. Lane C, sample at 72 h digested with Bgl II to show the linear DNA.

Article Snippet: To verify the nature of these slowly migrating molecules, TNAs extracted from the samples at 24 and 72 h were incubated with Taq DNA polymerase and analyzed by Southern blot (Fig. B).

Techniques: Transfection, Incubation

Journal: Malaria Journal

Article Title: Detection of knockdown resistance (kdr) mutations in Anopheles gambiae: a comparison of two new high-throughput assays with existing methods

doi: 10.1186/1475-2875-6-111

Figure Lengend Snippet: Examples of AS-PCR products for kdr -e and kdr -w genotyping . Gels A to D show examples of AS-PCR results using four different protocols, A [29], B [11], C [12], D [20]. Gels E to G show the result of using different DNA polymerases on the AS-PCR method described by [20], E: Promega PCR master mix, F: Qiagen HotStar Taq G:Finzymes Dynazyme II Gel H is an example of results of using the protocol of [20] with the Promega PCR master mix to genotype samples using the kdr -e assay from the 96 reference plate and gel I for the kdr -w assay. The same DNA templates were used in PCRs shown in gels A to G and from left to right were 100 bp DNA Ladder (Fermentas), homozygous wildtype, homozygous wildtype, heterozygous, heterozygous, homozygous mutant, homozygous mutant.

Article Snippet: PCR was carried out in a 25 μl volume with a final concentration of 1 × Buffer, 2 mM MgCl2 , 0.2 mM dNTPs, 0.1 μM each primer, 0.034 U/μl Taq DNA polymerase (Qiagen).

Techniques: Polymerase Chain Reaction, Mutagenesis

Journal: PLoS ONE

Article Title: Sex Determination in Highly Fragmented Human DNA by High-Resolution Melting (HRM) Analysis

doi: 10.1371/journal.pone.0104629

Figure Lengend Snippet: Melting temperature curve profiles produced by male and female samples in a HRM analysis. HRM analysis starts with a PCR amplification of the amelogenin gen. The amplification is in the presence of EvaGreen a saturating dye which presents high fluorescence when bound to dsDNA. Amplification is followed by a High Resolution Melting analysis (HRM) in the LightCycler 480 Real-Time PCR Instrument (Roche Applied Science). A dissociation curve analysis displays the different melting temperature of products from the X and Y chromosome.

Article Snippet: Lightcycler Master-Mix [composed by FastStart Taq DNA Polymerase, reaction buffer, dNTP mix (with dUTP instead of dTTP) and High Resolution Melting Dye] and nuclease-free water (QIAGEN), and 0.2 µM of each of the two primers: Amel_F ( 5′-CCCTTTGAAGTGGTACCAGAG-3′ ) and Amel_R (5′GGGAATAARGAACAAAATGTCTAC-3′, R = A or G).

Techniques: Produced, Polymerase Chain Reaction, Amplification, Fluorescence, Real-time Polymerase Chain Reaction

Journal: Indian Journal of Microbiology

Article Title: Simultaneous Detection of Major Pome Fruit Viruses and a Viroid

doi: 10.1007/s12088-013-0431-y

Figure Lengend Snippet: Sensitivity limits of multiplex RT-PCR. Lane 1 Amplification from tenfold diluted cDNA; lanes 2–6 amplicons from 10 −2 to 10 −6 fold diluted cDNA. Lanes 1–6 amplicons obtained with multiplexing kit; lanes 7–12 amplification with hot-start Taq DNA polymerase at same dilutions; lane M 100 bp marker

Article Snippet: The multiplex RT-PCR assay was optimized with regular Taq (Bangalore Genei, Bengaluru, India), Hot Start Taq (Fermentas, Vilnus, Lithuania) DNA polymerase and multiplexing kit from Qiagen under the conditions described above with nad5 as an internal control.

Techniques: Multiplex Assay, Reverse Transcription Polymerase Chain Reaction, Amplification, Multiplexing, Marker