phusion buffer gc (New England Biolabs)

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Name:

Phusion GC Buffer Pack

Description:

Phusion GC Buffer Pack 6 0 ml

Catalog Number:

b0519s

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Size:

6 0 ml

Category:

Buffers

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Structured Review

New England Biolabs phusion buffer gc

Phusion GC Buffer Pack 6 0 ml
https://www.bioz.com/result/phusion buffer gc/product/New England Biolabs
Average 99 stars, based on 198 article reviews
Price from $9.99 to $1999.99

phusion buffer gc - by Bioz Stars, 2020-09

99/100 stars

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Images

1) Product Images from "High-throughput mutagenesis using a two-fragment PCR approach"

Article Title: High-throughput mutagenesis using a two-fragment PCR approach

Journal: Scientific Reports

doi: 10.1038/s41598-017-07010-4

Agarose gel electrophoresis analysis of PCR fragments multiplied by Phusion High-Fidelity PCR Master Mix with HF Buffer and Phusion High-Fidelity PCR Master Mix with GC Buffer. Analysed PCRs are labelled as the mutated residue and the letter indicating whether the fragment is multiplied by mutation-specific forward (F) or reverse (R) primer. Expected PCR fragments are between 3000 and 4000 bp long. While using the PCR master mix with GC buffer gave expected fragments in all shown cases, using the Phusion High-Fidelity polymerase with HF buffer failed to multiply four fragments.

Figure Legend Snippet: Agarose gel electrophoresis analysis of PCR fragments multiplied by Phusion High-Fidelity PCR Master Mix with HF Buffer and Phusion High-Fidelity PCR Master Mix with GC Buffer. Analysed PCRs are labelled as the mutated residue and the letter indicating whether the fragment is multiplied by mutation-specific forward (F) or reverse (R) primer. Expected PCR fragments are between 3000 and 4000 bp long. While using the PCR master mix with GC buffer gave expected fragments in all shown cases, using the Phusion High-Fidelity polymerase with HF buffer failed to multiply four fragments.

Techniques Used: Agarose Gel Electrophoresis, Polymerase Chain Reaction, Mutagenesis

2) Product Images from "Solid-phase cloning for high-throughput assembly of single and multiple DNA parts"

Article Title: Solid-phase cloning for high-throughput assembly of single and multiple DNA parts

Journal: Nucleic Acids Research

doi: 10.1093/nar/gkv036

Results from head-to-tail SPC. ( A ) Comparison of compatibility of thermostable polymerases with the head-to-tail method. The polymerases with best proof-reading capabilities, Phusion and Deep Vent, all failed to generate transformants with the standard head-to-tail protocol. ( B ) Activity was retained for protocols using Phusion after supplementation of the washing buffer with SDS. ( C ) Colony screens of inserted region representing a selection of assemblies of various lengths and number of inserts assembled by head-to-tail SPC. Final construct sizes spanned 2.9 to 7.2 kbps.

Figure Legend Snippet: Results from head-to-tail SPC. ( A ) Comparison of compatibility of thermostable polymerases with the head-to-tail method. The polymerases with best proof-reading capabilities, Phusion and Deep Vent, all failed to generate transformants with the standard head-to-tail protocol. ( B ) Activity was retained for protocols using Phusion after supplementation of the washing buffer with SDS. ( C ) Colony screens of inserted region representing a selection of assemblies of various lengths and number of inserts assembled by head-to-tail SPC. Final construct sizes spanned 2.9 to 7.2 kbps.

Techniques Used: Activity Assay, Selection, Construct

3) Product Images from "Restriction enzyme-free mutagenesis via the light regulation of DNA polymerization"

Article Title: Restriction enzyme-free mutagenesis via the light regulation of DNA polymerization

Journal: Nucleic Acids Research

doi: 10.1093/nar/gkp150

Effects of a caged thymidine nucleobase on the PCR. ( A ) PCR product using Phusion DNA Polymerase. Polymerization is halted in the presence of a caging group, which is confirmed using the truncated primer P3 that affords the same length product. ( B ) PCR product using Taq DNA polymerase. Polymerization is halted in the presence of a caging group, which is confirmed using the truncated primer P3 that affords the same length product.

Figure Legend Snippet: Effects of a caged thymidine nucleobase on the PCR. ( A ) PCR product using Phusion DNA Polymerase. Polymerization is halted in the presence of a caging group, which is confirmed using the truncated primer P3 that affords the same length product. ( B ) PCR product using Taq DNA polymerase. Polymerization is halted in the presence of a caging group, which is confirmed using the truncated primer P3 that affords the same length product.

Techniques Used: Polymerase Chain Reaction

Polymerase Chain Reaction:

Article Title: Genomic and Genetic Variation in E2F1 in Men with Non-Obstructive Azoospermia
Article Snippet: .. For PCR, 50ng of gDNA was amplified using Phusion High-Fidelity PCR Master Mix with GC Buffer (New England Biolabs, MA). .. PCR products were purified using the ExoSAP-It kit (USB Scientific, OH) and sequenced on ABI 3730 x lDNA analyzers with capillary electrophoresis and fluorescent dye terminator detection (Genewiz Inc., NJ).

Article Title: Strand-specific RNA sequencing reveals extensive regulated long antisense transcripts that are conserved across yeast species
Article Snippet: .. Third, PCR was performed with Phusion High-Fidelity DNA Polymerase with GC buffer (New England Biolabs) and 2 M betaine (Sigma). .. Fourth, PCR primers were removed using 1.8× volume of AMPure PCR Purification kit (Beckman Coulter Genomics - Danvers, MA, USA).

Article Title: Restriction enzyme-free mutagenesis via the light regulation of DNA polymerization
Article Snippet: .. The forward and reverse primers (5 μl each of a 10 μM solution), the pGFPuv template (1 μl of a 0.1 ng/μl solution), dNTPs (2 μl each of a 1 mM solution), Phusion GC Buffer (5 μl of a 10× stock, New England Biolabs), Phusion DNA Polymerase (1 μl, two units) and dH2 O (31 μl) were mixed and subjected to the following PCR program: 95°C (2 min), followed by 40 cycles of 95°C (30 s), 40°C (60 s) and 72°C (3.3 min), with a final extension at 72°C (2 min). .. An identical PCR reaction was then repeated, using 5 μl of the previous reaction as the template, followed by purification with a PCR cleanup kit (Promega).

Article Title: Composition and Genetic Diversity of the Nicotiana tabacum Microbiome in Different Topographic Areas and Growth Periods
Article Snippet: .. The PCR reactions were carRIed out in 20 μL reaction mixtures, COntaining 5.8 μL sterile distillED water, 10.0 μL 5× Phusion® High-Fidelity PCR Master Mix with GC Buffer (New England Biolabs, Ipswich, Ma, USA), 1.0 μL forwARd primeR (10 μM), 1.0 μL reveRSe primeR (10 μM), 0.2 μL Taq DNA PolymERase, and 2.0 μL template DNA. .. The PCR amplification conditions were as follows: initial denaturation at 95 °C for 2 min, 30 cycles of denaturation at 95 °C for 30 s, annealing at 50 °C for 20 s, elongation at 72 °C for 60 s, elongation at 72 °C for 10 min, and then held at 4 °C.

Article Title: Methanogens and Iron-Reducing Bacteria: the Overlooked Members of Mercury-Methylating Microbial Communities in Boreal Lakes
Article Snippet: .. For this PCR amplification targeting the hgcA gene , we used 50 μl master mix containing 1× Phusion GC buffer, 0.2 mM dNTP mix, 5% dimethyl sulfoxide (DMSO), 0.1 μM each primer with generic adaptors, 7 μg/μl BSA, 4 μl extracted DNA template, and 1.0 U Phusion high-fidelity DNA polymerase (NEB, UK). .. The PCR program started with an initial 2-min denaturation at 98°C followed by 35 amplification cycles (10 s at 96°C, 30 s 56.5°C, and 45 s at 72°C), and a final 7-min extension at 72°C.

Article Title: High-throughput mutagenesis using a two-fragment PCR approach
Article Snippet: .. For truncations of the AtETR1 and LeETR1 plasmids by the two-fragment approach, a slightly different PCR protocol was used: 20 μL PCR mixture prepared with nuclease-free water (Cell Signaling Technology) contained 1 ng DNA template, 500 nM each primer, 200 μM each dNTP (deoxynucleoside triphosphate; NEB), 0.4 U μL−1 Phusion High-Fidelity DNA Polymerase (NEB), and either 1× Phusion buffer HF (NEB) or 3% (v/v) DMSO (NEB) with 1× Phusion buffer GC (NEB). .. Also, PCR thermocycling described above was modified in case of the ETR1 plasmids to have: (1) 11 step-down cycles with annealing starting from 65 °C down to 60 °C (i.e. decreasing by 0.5 °C in each subsequent cycle); (2) elongation time of at least 30 s per kbp of the target product (up to 200 s for the longest PCR products); and (3) final elongation time of 5 min.

Article Title: Changes in the intestinal microecology induced by bacillus subtilis inhibit the occurrence of ulcerative colitis and associated cancers: a study on the mechanisms
Article Snippet: .. Amplification of variable V4 region of 16S rRNA: The variable V4 region was amplified with Phusion® High-Fidelity PCR Master Mix with GC Buffer (New England Biolabs) and efficient and high-fidelity enzymes. .. The diluted DNA served as a template, and 515F-806R primers with Barcode were used for PCR.

Amplification:

Hybridization:

Article Title: Solid-phase cloning for high-throughput assembly of single and multiple DNA parts
Article Snippet: .. Extension An extension mix containing 17.25 μl water, 2.5 μl Phusion buffer (10×), 2.5 μl dNTPs (2 mM) and 0.25 μl Phusion (2 U/ μl New England Biolabs, Ipswich, MA, USA) was pre-heated at 65°C before used to resuspend the beads from the hybridization. .. Phusion HF was used as preferred buffer for extension unless the insert was PCR amplified using a different buffer; in those instances the matching buffer (e.g. Phusion GC buffer) was used also during the extension.