Top Ten Peer-Reviewed Articles
From NGS library construction to optimal sorting of mitotic cells, BioTechniques has presented you with ample new methods to improve your research this year. Here we list our ten most popular peer-reviewed papers, based on downloads, from the first half of 2014.
construction for next-generation sequencing: Overviews and challenges
In our first Practical Guide article of 2014, Head et al. review the
critical role of the sequencing library and assess the challenges
that may arise during its creation. And of course, they also
recommend options for troubleshooting and resolving these problems.
a BLAST-based tool for taxonomic classification of nucleotide
The microbiome is a hot topic these days, and taxonomic classification is an
important step in these studies. Here, Tuzhikov et al. present an open
source command line tool for microbial taxonomy analysis that
returns results even for sequences as short as 125 bp.
ligand-based protein quantification using immuno-PCR: A critical
review of single-probe and proximity ligation assays
qPCR has revolutionized gene expression analysis, but many researchers also
want to address protein translational patterns in their studies. In
this Practical Guide, Hansen et al. review a method that meets
this need: PCR-coupled protein quantification, often denoted as
immuno-PCR (iPCR), which targets soluble proteins.
amplification of target prior to PCR for improved low template DNA
Forensic DNA studies are often hampered by low quantities of poor quality DNA.
Here, Grisedale et al. present an amplification method for target
DNA prior to short tandem repeat (STR) analysis without introducing
in situ proximity (ChrISP): Single-cell analysis of chromatin
proximities at a high resolution
To overcome poor resolution or the need for a large number of cells, Chen et
al. combined the in situ proximity ligation technique with
conventional DNA FISH/immunostaining to create a new method for
looking at the proximity of different regions within chromatin.
of an integrated antibiotic resistance gene in mammalian cells to
re-enable antibiotic selection
2014 has brought several new
uses for the CRISPR/Cas9 system. Here, Ni et al. used the
endonuclease to disrupt antibiotic resistance genes in mammalian
cells, enabling researchers to perform additional rounds
of genetic manipulation with selection using the same
analysis of colony morphology in yeast
The structures of biofilms and microbial colonies can influence an organism's
phenotype. Here Ruusuvuori et al. present a new automated platform
for analyzing yeast colony morphology, offering an alternative to
subjective scoring of colonies and facilitating high-throughput
selection of modified aptamer pairs for diagnostic sandwich assays
Many techniques require pairs of analyte specific reagents for detection and
capture. Here, Ochsner et al. present a new method for isolating
aptamers and a screening method for determining the best pairs for
of using the QIAshredder instead of restriction digestion to prepare
DNA for droplet digital PCR
Genomic DNA can be difficult to partition into droplets or wells for digital
PCR. To overcome this, researchers use restriction enzymes to cut the
DNA into more manageable fragments. But here, Yukl et al. show that
using a QIAshredder spin column is quicker and easier. Not only that,
but it produces much more uniform fragments and avoids using
digestion buffers that can inhibit PCR.
analysis of mitosis-specific antibodies for bulk purification of mitotic
populations by fluorescence-activated cell sorting
Chromatin changes during mitosis have caught the attention of many biologists,
but to study these cells, it is necessary to first collect a lot of
them. Here, Campbell et al. present an optimized FACS-based method
for isolating fixed mitotic cells at virtually 100% mitotic purity.
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