Posts
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Rolling Circle Amplification (RCA) vs Bridge Amplification
Rolling Circle Amplification (RCA) and Bridge Amplification are two distinct amplification techniques used in molecular biology for DNA amplification. Each method has its own principles, applications, and advantages. Here’s a detailed comparison of the two: Rolling Circle Amplification (RCA) Definition: Rolling Circle Amplification is a method that amplifies circular DNA templates to produce long, repetitive…
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What are Read 1 and Read 2 in paired end sequencing?
In the context of DNA sequencing, Read 1 and Read 2 refer to the two sequencing reads generated from a DNA fragment during the sequencing process. Here’s a breakdown of each: Read 1: – Definition: Read 1 is the first sequence obtained from a DNA fragment during the sequencing process. – Process: During sequencing, the…
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What is MDA (Multiple Displacement Amplification)?
MDA stands for Multiple Displacement Amplification. It is a technique used to amplify DNA, particularly useful in scenarios where the starting material is limited, such as in single-cell genomics or when working with degraded samples. Significance of MDA: 1. High Sensitivity: MDA can amplify minute amounts of DNA, making it ideal for applications where only…
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Key sequencing steps in MGISEQ
The sequencing principle involves several key components, each playing a crucial role in the overall process. Here are the significant components and their functions: 1. DNA Nanoballs (DNBs): These are formed during the library preparation and amplification process. DNBs consist of multiple copies of the same DNA fragment, which enhances the signal during sequencing by…
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Detailed Library Preparation Steps of MGISEQ
1. Fragmentation of DNA: The genomic DNA (gDNA) is first fragmented to obtain the desired size for sequencing. 2. End Repair and A-tailing: The fragmented DNA undergoes end repair to create blunt ends, followed by the addition of an ‘A’ base to the 3′ ends to facilitate adapter ligation. 3. Adapter Ligation: Specific adapters are…
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How to assess the accuracy of SNP detection with DNA sequencing?
Comparison with Reference Data The authors conditionally considered the Illumina HiSeq 2500 data as a standard reference to evaluate the accuracy of the MGISEQ-2000 data. This involved calculating “error rates” such as “False Positive” and “False Negative” rates in the MGISEQ-2000 dataset (E704-M) using the Illumina dataset (E704-I) as a benchmark . Variant Calling Analysis…
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MGISEQ-2000 vs Illumina HiSeq 2500 platforms
Duplication Rates The MGISEQ-2000 exhibited a lower duplication rate compared to the Illumina HiSeq 2500 when analyzing individual fastq files. The overall duplication rate for the Illumina platform was higher (12.26%) due to the merging of fastq files from two different flow cells, while the duplication rate for both platforms did not exceed 5-6% in…
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What is a reversible terminator in DNA sequencing?
To identify the sequence in DNA, each base in a DNA fragment or DNA nanoball needs to be identified step-by-step. First, a primer binds to the adaptor in the DNA strand, and the primer will be extended using an artificial dNTP rather than a natural dNTP. The artificial dNTP usually prevents further extension, so one…
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What is the DNA sequencing platform, MGI-seq or DNB-seq?
MGI-seq or DNB-seq is a next-generation sequencing (NGS) platform developed by MGI Tech Co., Ltd. It could perform whole-genome sequencing (WGS), sequencing of cell-free DNA fragments (cfDNA), ATAC-Seq, etc. It is also a form of sequencing by synthesis (SBS) technology. But different from Illumina sequencing which utilizes bridge amplification, DNB-Seq requires the rolling circle amplification…