Prism ready reaction dye terminator cycle sequencing kit

Manufactured by Thermo Fisher Scientific

Sourced in United States

The PRISM™ Ready Reaction Dye Terminator Cycle Sequencing Kit is a pre-mixed reagent solution for DNA sequencing using the Sanger method. It contains the necessary reagents, including fluorescently labeled dye terminators, to perform the DNA sequencing reaction in a single tube.

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Lab products found in correlation

Abi prism 3500xl dna sequencer, by Thermo Fisher Scientific (2 mentions) Dreamtaq dna polymerase, by Thermo Fisher Scientific (1 mentions) Zr fungal bacterial dna kit, by Zymo Research (1 mentions)

Close spelling variants of this product

Ready Reaction Cycle Sequencing Kit (2 citations) Dye terminators (2 citations) ABI Prism® Big DyeTM Terminator v. 3.0 Ready Reaction Cycle Sequencing Kit (2 citations)

2 protocols using prism ready reaction dye terminator cycle sequencing kit

Sanger Sequencing of Purified PCR Products

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The sanger sequencing of the purified PCR products was done at Inqaba Biotechnical Industrial (Pty) Ltd, Pretoria, South Africa with PRISM™ Ready Reaction Dye Terminator Cycle sequencing kit using the dideoxy chain termination method and electrophoresed with a model ABI PRISM^® 3500XL DNA Sequencer (Applied Biosystems, Foster City, CA, USA) following manufacturer’s instructions.
For good quality sequence assurance, ChromasLite version 2.33 software was used for the analysis of chromatograms (sense and antisense) resulting from sequencing reaction (Technelysium 2004 ). The resulting chromatograms were edited using BioEdit Sequence Alignment Editor (Hall 2004 ). After this, the resulting consensus 16S rDNA sequences obtained were blasted in the NCBI (http://www.ncbi.nlm.nih.gov) database with the BLASTn for homology in order to identify the probable organism in question (Altshul et al. 1997 (link)). The sequences were deposited in the GenBank.

Adeniyi M., Titilawo Y., Oluduro A., Odeyemi O., Nakin M, & Okoh A.I. (2018). Molecular identification of some wild Nigerian mushrooms using internal transcribed spacer: polymerase chain reaction. AMB Express, 8, 148.

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Bacterial 16S rRNA Sequence Analysis

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Extraction of total DNA was achieved using the ZR Fungal/Bacterial DNA Kit™ (Zymo Research, Irvine, CA, USA) according to the manufacturer's instructions. Target 16S rRNA regions were amplified using DreamTaq™ DNA polymerase (Thermo Scientific™, Thermo Fisher Scientific, Waltham, MA, USA) with the bacterial universal primers 27F (5 -AGAGTTTGATCMTGGCTCAG-3 ) and 1492R (5 -CGGTTACCTTGTTACGACTT-3 ) as described by Lane et al. [42] and Turner et al. [43] .
Sanger sequencing of the purified PCR products was carried out by Inqaba Biotechnical Industries (Pty) Ltd., Hatfield, South Africa using the PRISM™ Ready Reaction Dye Terminator Cycle sequencing kit using chain termination by the dideoxy-nucleotides methodology and electrophoresed on an ABI PRISM ® 3500XL DNA Sequencer (Applied Biosystems, Foster City, CA, USA) following the manufacturer's instructions. Resultant chromatograms were analysed (Chromas version 2.6.6) as outlined in Technelysium [44] , followed by pairwise alignment in BioEdit Sequence Alignment Editor, version 7.0.5.3 [45] , and the resultant gene sequences compared with those already deposited (https://www.ncbi.nlm.nih.gov/).

Titilawo Y., Masudi W.L., Olawale J.T., Sekhohola-Dlamini L., & Cowan A.K. (2020). Coal-Degrading Bacteria Display Characteristics Typical of Plant Growth Promoting Rhizobacteria. Processes, 8(9), 1111-1111.

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