phusion (New England Biolabs)


Name:
Phusion GC Buffer Pack
Description:
Phusion GC Buffer Pack 6 0 ml
Catalog Number:
b0519s
Price:
24
Category:
Buffers
Size:
6 0 ml
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Structured Review

Phusion GC Buffer Pack 6 0 ml
https://www.bioz.com/result/phusion/product/New England Biolabs
Average 97 stars, based on 1 article reviews
Price from $9.99 to $1999.99
Images
1) Product Images from "RF-Cloning.org: an online tool for the design of restriction-free cloning projects"
Article Title: RF-Cloning.org: an online tool for the design of restriction-free cloning projects
Journal: Nucleic Acids Research
doi: 10.1093/nar/gks396

Figure Legend Snippet: RF-Cloning.org output page. (1) A unique 32 byte hash code is generated for all new projects, and is present in the URL for bookmarking purposes. (2) The hybrid primers are color coded, blue for sequence complementary to the plasmid, and green for the insert. The length of the primers can be adjusted by clicking on the arrow buttons if the user wishes to alter the annealing temperature. (3) If the insert site needs to be adjusted, the user can use the provided arrow buttons. (4) The secondary PCR conditions are optimized for iProof or Phusion as the polymerase, so the user should follow manufacturer’s instructions if using another high fidelity enzyme. ‘Insert’ refers to the mega-primer purified from the primary PCR reaction. (5) The entire sequence of the new plasmid is output, with insert in green and parental plasmid in blue. (6) The plasmid map can be drawn by specifying the positions of markers manually, or by auto-finding common features. Restriction enzyme cut sites can also be specified or automatically identified. If desired, the plasmid can be exported as a genbank file. (7) All projects are automatically saved, but making changes to the output page will activate the save button so those changes can be uploaded to the database. If the user has registered an account to access the plasmid management system, the save button will attach the project to their profile. (8) After the project has been completed and sent for sequencing, the sequencing results can be copied into a popup window for BLAST2 sequence alignment.
Techniques Used: Clone Assay, Polyacrylamide Gel Electrophoresis, Generated, Sequencing, Plasmid Preparation, Polymerase Chain Reaction, Purification
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

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 "In vitro synthesis of gene-length single-stranded DNA"
Article Title: In vitro synthesis of gene-length single-stranded DNA
Journal: Scientific Reports
doi: 10.1038/s41598-018-24677-5

Figure Legend Snippet: ssDNA production by aPCR. ( a ) aPCR reactions were assembled with a 50-molar excess of a forward primer for the amplification of a 1,000 nt ssDNA fragment using the M13mp18 ssDNA plasmid as template, and with 10 different polymerases that were tested for highest yield of ssDNA production (upper band: expected dsDNA size is 1,000 bp; lower band: expected ssDNA size is 1,000 nt) as judged by agarose gel electrophoresis (right panel). QuantaBio AccuStart HiFi, polymerase (lane 2, boxed) produced the highest amount without overlapping dsDNA contaminants. 1. Accustart; 2. Accustart HiFi; 3. Accustart II; 4. AccuPrime; 5. GoTaq; 6. DreamTaq; 7. Phusion; 8. Platinum SuperFi; 9. Q5; 10. Tth polymerase. ( b ) Biochemical validation of ssDNA production by incubating 1,000 nt aPCR reaction products with the ssDNA-specific ExoI or S1 nucleases or dsDNA-specific restriction enzymes Eco RI and Nae I (left panel). Agarose gel electrophoresis of the digestion products as labeled by lane (right panel). M: Marker, C: aPCR product control, ExoI: exonuclease I, S1: S1 nuclease, Enz: Eco RI + Nae I. ( c ) NEB LongAmp was used to generate ssDNA up to 15,000 nt long using lambda phage dsDNA as template. Purification of the 10 kb fragment shows a single band of higher molecular weight than the M13mp18 ssDNA (7,249 nt). ( d ) The primer design algorithm aPrime was used to select primers for product sizes between 500 and 3,000 nt using M13mp18 ssDNA as template and the Quantabio Accustart HiFi enzyme. SYBR Safe stained agarose gels illuminated under blue light show dsDNA as yellow bands, while ssDNA show as orange bands.
Techniques Used: Amplification, Plasmid Preparation, Agarose Gel Electrophoresis, Produced, Labeling, Marker, Purification, Molecular Weight, Staining
4) Product Images from "In situ 10-cell RNA sequencing in tissue and tumor biopsy samples"
Article Title: In situ 10-cell RNA sequencing in tissue and tumor biopsy samples
Journal: Scientific Reports
doi: 10.1038/s41598-019-41235-9

Figure Legend Snippet: A blend of Taq–Phusion polymerases improves selective poly(A) amplification of cDNA and reduces AL1 primer requirements. Cells were obtained by LCM from a human breast biopsy and split into 10-cell equivalent amplification replicates. ( A ) Poly(A) PCR was performed with 15 µg of AL1 primer with Taq alone (10 units), Phusion alone (4 units) or Taq/Phusion combination (3.75 units/1.5 units). ( B ) Poly(A) PCR was performed with either 25, 5, 2.5, or 0.5 µg of AL1 primer and the Taq–Phusion blend from (A). Above—Relative abundance for the indicated genes and preamplification conditions was measured by quantitative PCR (qPCR). Data are shown as the median inverse quantification cycle (40–Cq) ± range from n = 3 amplification replicates and were analysed by two-way (A) or one-way (B) ANOVA with replication. Below—Preamplifications were analysed by agarose gel electrophoresis to separate poly(A)-amplified cDNA from nonspecific, low molecular-weight concatemer (n.s.). Qualitatively similar results were obtained separately three times. Lanes were cropped by poly(A) PCR cycles for display but were electrophoresed on the same agarose gel and processed identically. The uncropped image is shown in Supplementary Fig. S13A .
Techniques Used: Amplification, Laser Capture Microdissection, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Agarose Gel Electrophoresis, Molecular Weight
Related Articles
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 Polymerase Chain Reaction:Article Title: Restriction enzyme-free mutagenesis via the light regulation of DNA polymerization Article Snippet: It is important to note that the selection of primers is key, and for some sequences, longer hybridization arms are required (see GFP deletion in ), which in turn requires additional caging groups to prevent undesirable hybridization during the PCR (see ‘Results and Discussion’ section). .. 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), Article Title: In situ 10-cell RNA sequencing in tissue and tumor biopsy samples Article Snippet: Poly(A) PCR Poly(A) PCR was carried out with several modifications to the earlier procedure , . .. To each tailed sample, 90 µl of poly(A) PCR buffer was added to a final concentration of 1x ThermoPol buffer (New England Biolabs), 2.5 mM MgSO4 , 1 mM dNTPs (Roche), 100 µg/ml BSA (Roche), 3.75 U Taq polymerase (NEB) and 1.5 U Article Title: RF-Cloning.org: an online tool for the design of restriction-free cloning projects Article Snippet: RECOMMENDED RF-CLONING PROTOCOL By default, the starting hybrid primers will be at least 40 bp long with an annealing temperature of at least 55°C for the primary PCR (amplification of insert) and at least 60°C for the secondary PCR (extension around the plasmid). .. High-fidelity DNA polymerase should be used for all PCR reactions, and it has been our experience that iProof (BioRad, Hercules, CA, USA) and Article Title: High-throughput mutagenesis using a two-fragment PCR approach Article Snippet: When using KOD Hot Start Master Mix, the following thermocycling setup was used: Initial denaturation at 95 °C for 2 min, followed by 20 step-down thermal cycles, each comprising denaturation at 95 °C for 20 s, annealing from 65 °C down to 55.5 °C for 10 s (0.5 °C decrement per cycle), and elongation at 70 °C for at the appropriate time (up to 135 s), then 5 thermal cycles with the constant annealing temperature (denaturation at 95 °C for 20 s, annealing at 62.5 °C for 10 s, elongation at 70 °C with the same time period as used in the step-down cycles), completed with a hold at 10 °C. .. 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× Article Title: Studying safe storage time of orange peel (Citrus reticulata) using high‐throughput sequencing and conventional pure culture, et al. 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Shimadzu; DM4000M microscope (Leica, Frankfurt, Germany); TruSeq® DNA PCR‐Free Sample Preparation Kit (Illumina Co., San Diego, CA, USA); QIAquick gum recovery kit (QIAGEN, Duesseldorf, Germany); Phusion® High‐Fidelity PCR Master Mix with Concentration Assay:Article Title: In situ 10-cell RNA sequencing in tissue and tumor biopsy samples Article Snippet: Poly(A) PCR Poly(A) PCR was carried out with several modifications to the earlier procedure , . .. To each tailed sample, 90 µl of poly(A) PCR buffer was added to a final concentration of 1x ThermoPol buffer (New England Biolabs), 2.5 mM MgSO4 , 1 mM dNTPs (Roche), 100 µg/ml BSA (Roche), 3.75 U Taq polymerase (NEB) and 1.5 U Microscopy:Article Title: Studying safe storage time of orange peel (Citrus reticulata) using high‐throughput sequencing and conventional pure culture, et al. Studying safe storage time of orange peel (Citrus reticulata) using high‐throughput sequencing and conventional pure culture Article Snippet: Retsch MM400 Ball Mill (Laichi co., Berlin, Germany); PTC200PCR (Bio‐Rad, Berkeley, CA, USA); GelDox XR gel imaging system (Bio‐Rad, USA); DYY‐8C Electrophoretic apparatus (Beijing Liuyi Instrument, China); BI3730XL sequenator (Applied Biosystems, Foster City, CA, USA); KRQ‐300P manual climatic box (ChongQing YinHe Test Instrument, China); Plant DNA Extraction Kit (TIANGEN Biotech Co., Chengdu, China); 2 × Taq PCR Master Mix (TIANGEN Biotech Co.,); primer was compounded by Sangon Co., China. .. Shimadzu; DM4000M microscope (Leica, Frankfurt, Germany); TruSeq® DNA PCR‐Free Sample Preparation Kit (Illumina Co., San Diego, CA, USA); QIAquick gum recovery kit (QIAGEN, Duesseldorf, Germany); Phusion® High‐Fidelity PCR Master Mix with Sample Prep:Article Title: Studying safe storage time of orange peel (Citrus reticulata) using high‐throughput sequencing and conventional pure culture, et al. Studying safe storage time of orange peel (Citrus reticulata) using high‐throughput sequencing and conventional pure culture Article Snippet: Retsch MM400 Ball Mill (Laichi co., Berlin, Germany); PTC200PCR (Bio‐Rad, Berkeley, CA, USA); GelDox XR gel imaging system (Bio‐Rad, USA); DYY‐8C Electrophoretic apparatus (Beijing Liuyi Instrument, China); BI3730XL sequenator (Applied Biosystems, Foster City, CA, USA); KRQ‐300P manual climatic box (ChongQing YinHe Test Instrument, China); Plant DNA Extraction Kit (TIANGEN Biotech Co., Chengdu, China); 2 × Taq PCR Master Mix (TIANGEN Biotech Co.,); primer was compounded by Sangon Co., China. .. Shimadzu; DM4000M microscope (Leica, Frankfurt, Germany); TruSeq® DNA PCR‐Free Sample Preparation Kit (Illumina Co., San Diego, CA, USA); QIAquick gum recovery kit (QIAGEN, Duesseldorf, Germany); Phusion® High‐Fidelity PCR Master Mix with |