• Volume 10, 2020
  • Volume 9, 2019
  • Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011

Developmental Biology

Tandem Affinity Purification in Drosophila Heads and Ovaries Authors:  Anita Pepper, Balpreet Bhogal and Thomas Jongens, date: 08/05/2012, view: 11621, Q&A: 0
Tandem affinity purification (TAP) (Pugi et al.,2001; Rigaut et al., 1999) is a method that uses a tagging approach of a target protein of interest for a two-step purification scheme in order to pull down protein complexes under native conditions and expression levels. The TAP tag consists of three components: a ...
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Microbiology

Construction and Screening of a Transposon Insertion Library of Yersinia enterocolitica (YeO3-R1) Authors:  Maria Pajunen, Elise Pinta and Mikael Skurnik, date: 08/05/2012, view: 11950, Q&A: 0
The Mu-transposon system is one of the best characterized transposition systems. Under minimal in vitro set-up, Mu transposition requires only a simple reaction buffer, MuA transposase protein, mini-Mu transposon DNA (donor) and target DNA. The reaction proceeds via initial assembly of the transposition complex that directs transposon ...
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Molecular Biology

5-Hydroxymethylcytosine (5-hmC) Specific Enrichment Authors:  Keith E Szulwach, Chun-Xiao Song, Chuan He and Peng Jin, date: 08/05/2012, view: 8210, Q&A: 0
5-Hydroxymethylcytosine (5-hmC) is a newly discovered DNA modification in mammalian genomes. This protocol is to be a highly efficient and selective chemical approach to label and capture 5-hmC, taking advantage of a bacteriophage enzyme that adds a glucose moiety to 5-hmC specifically, which could in turn be used for high-throughput mapping via ...
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Stable Interference by shRNA with pSUPER.retro Vectors and Lipofectamine Author:  Silvia Soddu, date: 08/05/2012, view: 11359, Q&A: 0
RNA interference is a powerful genetic approach for efficiently silencing target genes. Expression of short hairpin RNAs (shRNAs) allows analysis of the consequences of stably silencing genes. This protocol describes a method to stably integrate shRNA constructs with pSUPER.retro vectors and Lipofectamine in RKO and H1299 cells. This method can be ...
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A Quick, No Frills Approach to Mouse Genotyping Author:  Manuel E. Lopez, date: 08/05/2012, view: 22115, Q&A: 1
Mice are extremely powerful mammalian genetic model organisms for basic and medical research, but managing a colony of transgenic mice is time consuming and expensive, many times requiring the help of dedicated technicians. Slow and laborious genotyping procedures add to the hassle. Outsourcing is costly and may not be as fast as desired, ...
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