The schematic is the same, and part numbers are the same. The differences between the 1.5 and 1.6 are fairly minor - the circuit is identical to the previous version, all the parts other than some chassis mounted components are the same. To ensure maximum yields from minimum amounts of starting material, an optional high-fidelity amplification step can be performed.This is the build guide for the Amp Camp Amp v1.6. Fragments are then size selected and purified. Barcode adapters, which contain a unique identifying sequence, are also available with the GeneRead Library Prep (L) Kit and enable multiplex sequencing reactions to be performed. Afterwards, adaptors, which are necessary for amplification and sequencing, are ligated to both ends of the DNA fragments. Following fragmentation, the ends of the DNA fragments are repaired. Procedure for generating Ion Torrent-compatible DNA libraries Samples consisting of longer DNA fragments are first sheared into a random library of fragments that are a median fragment size of 400 bp (when using the Ion Torrent PGM instrument), or 200 bp (when using the Ion Proton instrument), in length. To ensure maximum yields from minimum amounts of starting material, an optional high-fidelity amplification step can also be performed. 180514), which allows precise, column-based size selection of DNA (see figure Precise size selection). Cleanup and size selection can be done using the GeneRead Size Selection Kit (cat. Barcode adapters, which contain a unique identifying sequence, are also available with the GeneRead Library Prep (I) Kit and enable multiplex sequencing reactions to be performed. Following fragmentation, the ends of the DNA fragments are repaired and an A-overhang is added at the 3'-end of each strand. Procedure for generating Illumina-compatible DNA libraries Samples consisting of longer DNA fragments are first sheared into a random library of fragments that are a median fragment size of 300 bp (when using the Illumina MiSeq instrument, version 1), or 500 bp (when using the Illumina MiSeq instrument, version 2), in length. The GeneRead HiFi Polymerase, together with its unique buffer formulation, ensures uniform amplification of genomic regions that contain highly variable GC content, thereby ensuring even coverage in subsequent sequencing reactions (see figure Better sequence coverage). Lower sequence bias when amplifying GC- and AT-rich regions In standard PCR amplification procedures, regions of DNA with high AT or GC content can result in little or no amplification, leading to misleading sequence data and NGS results. GeneRead HiFi Polymerase is a unique, highly specific and processive enzyme that delivers accurate amplification of library DNA with low error rates and minimum bias (see figure Low error rates with minimal sequence bias due to high-fidelity amplification). Low error rates with minimal sequence bias To ensure maximum yields from minimum amounts of starting material, GeneRead Library Prep Kits include an innovative high-fidelity master mix that includes GeneRead HiFi Polymerase for an optional amplification step. Highly efficient ligation reactions and an optional, high-fidelity amplification step ensure superior library yields with uniform coverage distribution, even from as low as 50 ng starting material (see figures High yields of library DNA with uniform coverage distribution and Superior yields of library DNA). High yields of library DNA with uniform coverage distribution GeneRead Library Prep Kits provide an efficient and optimized workflow to reproducibly generate high yields of DNA library, with minimal sequence bias and low error rates, for use on NGS platforms from Illumina or the Ion Torrent instrument from Life Technologies.
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