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Related Concept Videos

Single-Strand DNA Binding Proteins01:03

Single-Strand DNA Binding Proteins

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For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...
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Lagging Strand Synthesis01:59

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During replication, the complementary strands in double-stranded DNA are synthesized at different rates. Replication first begins on the leading strand. Replication starts later, occurs more slowly, and proceeds discontinuously on the lagging strand.
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Fixing Double-strand Breaks02:04

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The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...
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DNA Helicases00:55

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DNA unwinding helicase enzymes are a type of motor protein. Motor proteins can translocate along filaments or polymers using energy generated from ATP hydrolysis. Helicases are involved in all the important cellular processes where DNA unwinding is required, such as DNA replication, repair, recombination, and transcription. They are present in all living organisms, but vary in their structure, function, and mechanism of action. For example, in prokaryotes, DnaB helicase binds and translocates...
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Topoisomerases are enzymes that relax overwound DNA molecules during various cell processes, including DNA replication and transcription. These enzymes regulate positive and negative DNA supercoiling without changing the nucleotide sequence. DNA overwinding in a clockwise direction results in positively supercoiled DNA, whereas underwinding in a counterclockwise direction produces negatively supercoiled DNA.
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Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method
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Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method

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A Method for Single-Stranded Ancient DNA Library Preparation.

Marie-Theres Gansauge1, Matthias Meyer2

  • 1Department of Evolutionary Genetics, Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany. marie_gansauge@eva.mpg.de.

Methods in Molecular Biology (Clifton, N.J.)
|March 16, 2019
PubMed
Summary
This summary is machine-generated.

This study presents an improved genomic library preparation method using single DNA strands. This approach enhances efficiency when working with highly degraded DNA samples.

Keywords:
Ancient DNALibrary preparationNGSSingle-stranded ligationT4 DNA ligase

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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Genomic library preparation is crucial for DNA analysis.
  • Highly degraded DNA presents challenges for traditional library preparation methods.
  • Current methods can be inefficient with fragmented DNA, impacting downstream applications.

Purpose of the Study:

  • To develop a more efficient protocol for genomic library preparation from degraded DNA.
  • To optimize library construction using single DNA strands.
  • To improve the yield and quality of libraries derived from challenging DNA samples.

Main Methods:

  • A three-step protocol for single-stranded DNA library construction.
  • Utilizing T4 DNA ligase with a splinter oligonucleotide for first adapter ligation.
  • Employing a proofreading polymerase for DNA strand copying.
  • Performing blunt-end ligation of the second adapter to create double-stranded libraries.

Main Results:

  • Demonstrated increased efficiency in library preparation from degraded DNA.
  • Successfully constructed libraries from single DNA strands.
  • The protocol yields high-quality libraries suitable for various genomic analyses.
  • Showcased the effectiveness of the three-step single-strand approach.

Conclusions:

  • The described single-stranded library preparation protocol offers a significant improvement for degraded DNA.
  • This method enhances the efficiency and reliability of genomic library construction.
  • The protocol is a valuable tool for researchers working with low-quality or ancient DNA samples.