onetaq  (New England Biolabs)

Journal: bioRxiv

Article Title: Akaby - cell-free protein expression system for linear templates

doi: 10.1101/2021.11.03.467179

Figure Lengend Snippet: PCR amplified eGFP gene construct. The design of a 898 nt length DNA oligo. The oligo was PCR amplified from T7Max-GFP plasmid with Primer 11 and Primer 12 with LongAmp Taq 2X Master Mix (New England BioLabs Inc., M0287S). T7Max, the enhanced T7 polymerase promoter sequence; eGFP gene, an eGFP coding sequence; His, a 8xHistidine taq sequence; T500, a transcription terminator sequence.

Article Snippet: 10 μl total volume of colony PCR was performed with 0.4 μM of locus-(Primer 1 and 4) and insert-(Primer 2 and 3) specific primer pairs, 1X OneTaq® Quick-Load® 2X Master Mix with Standard Buffer (New England BioLabs Inc., M0486L), and 2 μl of water suspended colony.

Techniques: Polymerase Chain Reaction, Amplification, Construct, Plasmid Preparation, Sequencing

Journal: bioRxiv

Article Title: Akaby - cell-free protein expression system for linear templates

doi: 10.1101/2021.11.03.467179

Figure Lengend Snippet: PCR amplified eGFP gene construct. The design of a 898 nt length DNA oligo. The oligo was PCR amplified from T7Max-GFP plasmid with Primer 11 and Primer 12 with LongAmp Taq 2X Master Mix (New England BioLabs Inc., M0287S). T7Max, the enhanced T7 polymerase promoter sequence; eGFP gene, an eGFP coding sequence; His, a 8xHistidine taq sequence; T500, a transcription terminator sequence.

Article Snippet: The oligo was PCR amplified with OneTaq® 2X Master Mix with Standard Buffer (New England BioLabs Inc., M0482L), followed by agarose gel purification with GenCatch Advanced Gel Extraction Kit (Epoch Life Science, Inc., No. 2260050).

Techniques: Polymerase Chain Reaction, Amplification, Construct, Plasmid Preparation, Sequencing

Journal: bioRxiv

Article Title: The Macerprep: a minimalist kit- and enzyme-free high-yield miniprep utilising alkaline lysis and alkaline hydrolysis principles

doi: 10.1101/2020.08.13.249607

Figure Lengend Snippet: Macerprepped DNA is good template for PCR. 1-3, PCR product by NEB Q5® HiFi polymerase. 4-6, PCR product by NEB Onetaq polymerase. I, 4, Miraprepped DNA. 2, 5, Macerprepped DNA with Solution 3 process. 3, 6, Macerprepped DNA with Solution N3 process. M, marker.

Article Snippet: According to the NEB calculator (NEB, online tool), Tm for NEB Q5 and NEB Onetaq was determined to be 52°C and 61°C.

Techniques: Polymerase Chain Reaction, Marker

Journal: bioRxiv

Article Title: Akaby - cell-free protein expression system for linear templates

doi: 10.1101/2021.11.03.467179

Figure Lengend Snippet: PCR amplified eGFP gene construct. The design of a 898 nt length DNA oligo. The oligo was PCR amplified from T7Max-GFP plasmid with Primer 11 and Primer 12 with LongAmp Taq 2X Master Mix (New England BioLabs Inc., M0287S). T7Max, the enhanced T7 polymerase promoter sequence; eGFP gene, an eGFP coding sequence; His, a 8xHistidine taq sequence; T500, a transcription terminator sequence.

Article Snippet: The quantitative PCR reaction mix was prepared by mixing 2 μl of complementary DNA from the reverse transcription with 2 μl of 10 μM forward and reverse primers, 11.25 μl OneTaq Hot Start 2X Master Mix with Standard Buffer (Catalog No. M0484, New England BioLabs Inc.), 1.25 μl Chai Green Dye 20X (Catalog No. R01200, Chai Bio), and 7.5 μl of nuclease-free water.

Techniques: Polymerase Chain Reaction, Amplification, Construct, Plasmid Preparation, Sequencing