Purification of Oligonucleotides Using Denaturing Polyacrylamide Gel Electrophoresis

  title={Purification of Oligonucleotides Using Denaturing Polyacrylamide Gel Electrophoresis},
  author={Andrew D. Ellington and Jack D. Pollard},
  journal={Current Protocols in Molecular Biology},
Cloning vectors derived from filamentous phage are extremely useful because they allow cloned DNA to be isolated as either single‐ or double‐stranded DNA. This unit contains protocols for preparing both forms of DNA and for characterizing inserts in M13‐derived vectors. A protocol is also presented for preparing single‐stranded DNA from plasmids using superinfection with helper phage. This method is advantageous because it allows cloned DNA to be maintained in the form of a plasmid while… 

Production of Homogeneous Recombinant RNA Using a tRNA Scaffold and Hammerhead Ribozymes.

An efficient and facile method to release the RNA of interest from the tRNA scaffold by selective cleavage using cis-acting hammerhead ribozymes is described.

Liquid Chromatography‐Mass Spectrometry Analysis of DNA Polymerase Reaction Products

This unit describes experimental and analytical procedures for characterizing the efficiency and fidelity of translesion DNA synthesis across various DNA damages by DNA polymerases in vitro. This

Polyacrylamide Gel Electrophoresis (PAGE) of Synthetic Nucleic Acids

Protocols are given for analysis of oligon nucleotides by PAGE, using either methylene blue staining or radiolabeling to mark the oligonucleotide, and for purification by PAGE.

Enzymatic Synthesis of Backbone‐Modified Oligonucleotides Using T4 DNA Ligase

A small‐scale experiment is described to probe the efficiency of the ligation reaction of modified oligonucleotides in the presence of 3 M betaine and 10% PEG 8000, followed by large‐scale ligation with subsequent isolation of the ligated oligon nucleotide.

Steady‐State Kinetic Analysis of DNA Polymerase Single‐Nucleotide Incorporation Products

Data analysis procedures and fitting to steady‐state kinetic models is presented to highlight the kinetic differences involved in the bypass of damaged versus undamaged DNA.

Current Protocols in Chemical Biology Xenobiotic Nucleic Acid (XNA) synthesis by Phi29 DNA Polymerase ms#CP-17-0313-R1

Phi29 DNA Polymerase is presented as an efficient catalyst for the production of various artificial nucleic acids (XNAs) carrying backbone modifications such as 1, 5-anhydrohexitol nucleic acid (HNA), 2’-deoxy-2‘-fluoro3 arabinonucleic Acid (FANA), and 2′- fluoro-2′- deoxyribonucleIC acid (2’fluoro-DNA).

Overview of Blotting

The utilization of the blotting technology over the last 30 years has been instrumental to the elucidation of many fundamental biological processes and holds promise for even greater discovery over the next 30 years.

Isolation and Cloning of Small RNAs from Virus‐Infected Plants

This unit describes an isotope‐free small‐RNA cloning procedure that utilizes unmodified smallRNAs and is routinely used to characterize small RNAs from various plant tissues.



Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresis.

We describe the use of polyacrylamide gel electrophoresis to estimate chain lengths of double- and single-stranded DNA molecules in the size range 20-1000 base pairs (or nucleotides). Double-stranded

Identification and suppression of secondary structures formed from deoxy-oligonucleotides during electrophoresis in denaturing polyacrylamide-gels.

Nature and stability of secondary structures formed from DNA-fragments during electrophoresis in urea containing polyacrylamide-gels were investigated. Duplices and especially hairpin loops were

Recovery of DNA from gels.

  • H. Smith
  • Biology
    Methods in enzymology
  • 1980

Evaluation and Purification of Synthetic Oligonucleotides

  • User Bulletin no . 13
  • 1984