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DNA library preparation
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Quantification of DNA sequence tags from engineered constructs such as plasmids, transposons, or other transgenes underlies many functional genomics measurements. Typically, such measurements rely on PCR followed by next-generation sequencing. However, PCR amplification can introduce significant quantitative error.
17p
vigalileogalilei
27-02-2022
12
1
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Current library preparation protocols for Illumina HiSeq and MiSeq DNA sequencers require ≥2 nM initial library for subsequent loading of denatured cDNA onto flow cells. Such amounts are not always attainable from samples having a relatively low DNA or RNA input; or those for which a limited number of PCR amplification cycles is preferred (less PCR bias and/or more even coverage).
10p
vibeauty
23-10-2021
11
1
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Next-generation sequencing (NGS) is fundamental to the current biological and biomedical research. Construction of sequencing library is a key step of NGS. Therefore, various library construction methods have been explored. However, the current methods are still limited by some shortcomings.
12p
vibeauty
23-10-2021
13
0
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Transposome-based technologies have enabled the streamlined production of sequencer-ready DNA libraries; However, current methods are highly sensitive to the amount and quality of input nucleic acid.
16p
vitzuyu2711
29-09-2021
8
1
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Originally developed for the genomic analysis of highly degraded ancient DNA, single-stranded DNA (ssDNA) library preparation methods are gaining popularity in the field of cfDNA analysis due to their efficiency and ability to convert short, fragmented DNA into sequencing libraries without altering DNA ends. However, current ssDNA methods are costly and time-consuming.
14p
vijeeni2711
24-07-2021
7
0
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Barcode multiplexing is a key strategy for sharing the rising capacity of next-generation sequencing devices: Synthetic DNA tags, called barcodes, are attached to natural DNA fragments within the library preparation procedure. Different libraries, can individually be labeled with barcodes for a joint sequencing procedure.
14p
vikentucky2711
26-11-2020
14
0
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PCR amplification is an important step in the preparation of DNA sequencing libraries prior to high-throughput sequencing. PCR amplification introduces redundant reads in the sequence data and estimating the PCR duplication rate is important to assess the frequency of such reads.
11p
vioklahoma2711
19-11-2020
10
2
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Noises and artifacts may arise in several steps of the next-generation sequencing (NGS) process. Recently, an NGS library preparation method called SMART, or Switching Mechanism At the 5′ end of the RNA Transcript, is introduced to prepare ChIP-seq (chromatin immunoprecipitation and deep sequencing) libraries from small amount of DNA material, using the DNA SMART ChIP-seq Kit.
13p
vicoachella2711
27-10-2020
8
0
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Standardized Nucleic Acid Quantification for SEQuencing (SNAQ-SEQ) is a novel method that utilizes synthetic DNA internal standards spiked into each sample prior to next generation sequencing (NGS) library preparation.
14p
vikuala271
13-06-2020
7
0
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: Fragmentation at random nucleotide locations is an essential process for preparation of DNA libraries to be used on massively parallel short-read DNA sequencing platforms. Although instruments for physical shearing, such as the Covaris S2 focused-ultrasonicator system.
13p
viuchiha2711
21-04-2020
6
0
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Molecular Cytogenetics The introduction of FISH methodologies in the late 1980s revolutionized the field of cytogenetics. In principle, FISH is similar to other DNA-DNA hybridization methodologies. The probe is labeled with a hapten, such as biotin or digoxigenin, to allow detection with a fluorophore (e.g., FITC or rhodamine). After the hybridization step, the specimen is counter-stained and the preparations are visualized with a fluorescence microscope.
5p
konheokonmummim
03-12-2010
63
3
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