Springer protocols feed by Biotechnology 
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This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.Measuring the Elasticity of Ribonucleotide(s)-Containing DNA Molecules Using AFM
Ribonucleotides, ribonucleoside monophosphates (rNMPs), have been revealed as possibly the most noncanonical nucleotides in genomic DNA. rNMPs, either not removed from Okazaki fragments during DNA replication or incorporated and scattered throughout the genome, pose a perturbation to the structure and a threat to the stability of DNA. The instability of DNA is mainly due to the extra 2′-hydroxyl (OH) group of rNMPs which give rise to local structural effects, which may disturb various molecular interactions in cells. As a result of these structural perturbations by rNMPs, the elastic properties of DNA are also affect...
Source: Springer protocols feed by Biotechnology - April 25, 2015 Category: Biotechnology Source Type: news
Silver Nanoclusters for RNA Nanotechnology: Steps Towards Visualization and Tracking of RNA Nanoparticle Assemblies
The growing interest in designing functionalized, RNA-based nanoparticles (NPs) for applications such as cancer therapeutics requires simple, efficient assembly assays. Common methods for tracking RNA assemblies such as native polyacrylamide gels and atomic force microscopy are often time-intensive and, therefore, undesirable. Here we describe a technique for rapid analysis of RNA NP assembly stages using the formation of fluorescent silver nanoclusters (Ag NCs). This method exploits the single-stranded specificity and sequence dependence of Ag NC formation to produce unique optical readouts for each stage of RNA NP assemb...
Source: Springer protocols feed by Biotechnology - April 25, 2015 Category: Biotechnology Source Type: news
Large Scale Purification of RNA Nanoparticles by Preparative Ultracentrifugation
Purification of large quantities of supramolecular RNA complexes is of paramount importance due to the large quantities of RNA needed and the purity requirements for in vitro and in vivo assays. Purification is generally carried out by liquid chromatography (HPLC), polyacrylamide gel electrophoresis (PAGE), or agarose gel electrophoresis (AGE). Here, we describe an efficient method for the large-scale purification of RNA prepared by in vitro transcription using T7 RNA polymerase by cesium chloride (CsCl) equilibrium density gradient ultracentrifugation and the large-scale purification of RNA nanoparticles by sucrose gradie...
Source: Springer protocols feed by Biotechnology - April 25, 2015 Category: Biotechnology Source Type: news
HPLC Purification of RNA Aptamers up to 59 Nucleotides with Single-Nucleotide Resolution
An RNA sample is usually heterogeneous. RNA heterogeneity refers to difference in length or size (i.e., number of nucleotides [nt]), sequence, or alternative but coexisting conformations. Separation and purification of RNA is generally required for investigating the structure and function of RNA, such as RNA catalysis and RNA structure determination by nuclear magnetic resonance or crystallography. Separation and purification of RNA is also required for using RNAs as functional probes and therapeutics as well as building blocks for RNA nanoparticles. Previously established protocols are limited in separating RNAs longer th...
Source: Springer protocols feed by Biotechnology - April 25, 2015 Category: Biotechnology Source Type: news
Using RNA Nanoparticles with Thermostable Motifs and Fluorogenic Modules for Real-Time Detection of RNA Folding and Turnover In Prokaryotic and Eukaryotic Cells
RNA nanotechnology is an emerging field at the interface of biochemistry and nanomaterials that shows immense promise for applications in nanomedicines, therapeutics and nanotechnology. Noncoding RNAs, such as siRNA, miRNA, ribozymes, and riboswitches, play important roles in the regulation of cellular processes. They carry out highly specific functions on a compact and efficient footprint. The properties of specificity and small size make them excellent modules in the construction of multifaceted RNA nanoparticles for targeted delivery and therapy. Biological activity of RNA molecules, however, relies on their proper fold...
Source: Springer protocols feed by Biotechnology - April 25, 2015 Category: Biotechnology Source Type: news
Methods and Assays for Specific Targeting and Delivery of RNA Nanoparticles to Cancer Metastases
In recent years, RNA nanotechnology has become increasingly attractive due to its potential for applications in nanomedicine. RNA nanotechnology refers to the design and synthesis of nanoparticles composed mainly of RNA via bottom-up self-assembly. RNA nanoparticle is a suitable candidate for targeted delivery of therapeutics to cancer cells due to its multivalency, which allows the combination of therapeutic, targeting, and detection moieties all into one nanoparticle. To date, a system capable of exclusively targeting metastatic cancers that have spread to distant organs or lymph nodes is in demand. In this chapter, we r...
Source: Springer protocols feed by Biotechnology - April 25, 2015 Category: Biotechnology Source Type: news
Freeze-Drying of Mammalian Sperm
Long-term preservation of mammalian sperm at suprazero temperatures is desired to save storage and space costs as well as to facilitate transport of preserved samples. This can be accomplished by the freeze-drying of sperm samples. Although freeze-drying results in immotile and membrane-compromised sperm, intracytoplasmic sperm injection (ICSI) can be used to introduce such an immotile sperm into an oocyte and thus start the fertilization process. So far, it has been shown that improved freeze-drying protocols preserve chromosomal integrity and oocyte-activating factor(s) at 4 °C for several years and at ambient temper...
Source: Springer protocols feed by Biotechnology - December 2, 2014 Category: Biotechnology Source Type: news
Freeze-Drying of Decellularized Heart Valve Tissues
Decellularized xeno-antigen-depleted porcine pulmonary heart valves tissues may be used as matrix implants for patients with malfunctioning heart valves. Decellularized tissues are biological scaffolds composed of extracellular matrix components. Biological scaffolds closely resemble properties of native tissue, but lack immunogenic factors of cellular components. Decellularized heart valve scaffolds need to be stored to be readily available whenever needed. Scaffolds can be stored at reduced supra-zero temperatures, cryopreserved or freeze-dried. The advantage of freeze-drying is that it allows long-term storage at room t...
Source: Springer protocols feed by Biotechnology - December 2, 2014 Category: Biotechnology Source Type: news
Cord Blood Clinical Processing, Cryopreservation, and Storage
Allogeneic umbilical cord blood (UCB) hematopoietic stem cell transplantation has become a crucial advancement in the treatment for a variety of diseases including hematopoietic and non-hematopoietic malignancies, BM failure syndromes, hemoglobinopathies, and metabolic and immunodeficiency disorders. It has been well documented that the success of UCB engraftment is tied to UCB banking processes, and now there are established guidelines for standardization of collection, banking, processing, and cryopreservation for unrelated UCB units with purpose of achieving consistent production of high quality placental and UCB units ...
Source: Springer protocols feed by Biotechnology - December 2, 2014 Category: Biotechnology Source Type: news
Directional Freezing for Large Volume Cryopreservation
Cryopreservation is currently the method of choice when it comes to long-term preservation of viable biological samples. The process, and consequently the volume of the sample, however, is limited by the ability to achieve homogenous and efficient heat removal. When this cannot be properly managed, ice crystals will grow uncontrollably resulting in extensive damage to the cryopreserved cells or tissues. Directional freezing is a technique that can be used to precisely control heat dissipation and ice crystal growth and morphology even when freezing large volumes. The technique has been used over the years to cryopreserve s...
Source: Springer protocols feed by Biotechnology - December 2, 2014 Category: Biotechnology Source Type: news
Vitrification of Heart Valve Tissues
Application of the original vitrification protocol used for pieces of heart valves to intact heart valves has evolved over time. Ice-free cryopreservation by Protocol 1 using VS55 is limited to small samples where relatively rapid cooling and warming rates are possible. VS55 cryopreservation typically provides extracellular matrix preservation with approximately 80 % cell viability and tissue function compared with fresh untreated tissues. In contrast, ice-free cryopreservation using VS83, Protocols 2 and 3, has several advantages over conventional cryopreservation methods and VS55 preservation, including long-term preserv...
Source: Springer protocols feed by Biotechnology - December 2, 2014 Category: Biotechnology Source Type: news
Cryopreservation of Plant Cell Lines
Plant cell cultures may consist of dedifferentiated cells as well as of cells showing embryogenic potential. They can be used for very different purposes in research and biotechnology as well as for plant propagation. For such cell cultures, cryopreservation is the only means for long-term preservation. Most of the different cryopreservation approaches, which are generally used for plant tissues, have also been applied to plant cell cultures; they include slow freezing, vitrification, and encapsulation/dehydration approaches. The controlled-rate slow freezing approach which is described here, however, remains to be the gol...
Source: Springer protocols feed by Biotechnology - December 2, 2014 Category: Biotechnology Source Type: news
Writing Standard Operating Procedures (SOPs) for Cryostorage Protocols: Using Shoot Meristem Cryopreservation as an Example
Standard operating procedures are a systematic way of making sure that biopreservation processes, tasks, protocols, and operations are correctly and consistently performed. They are the basic documents of biorepository quality management systems and are used in quality assurance, control, and improvement. Methodologies for constructing workflows and writing standard operating procedures and work instructions are described using a plant cryopreservation protocol as an example. This chapter is pertinent to other biopreservation sectors because how methods are written, interpreted, and implemented can affect the quality of st...
Source: Springer protocols feed by Biotechnology - December 2, 2014 Category: Biotechnology Source Type: news
Freeze-Drying of Proteins
Freeze-drying has become one of the most important processes for the preservation of biological products. This chapter provides protocols for freeze-drying of proteins and discusses the importance of formulation, cycle development, and validation. Specific formulations for stabilization of proteins are presented as well as advice on common problems with freeze-drying of proteins. (Source: Springer protocols feed by Biotechnology)
Source: Springer protocols feed by Biotechnology - December 2, 2014 Category: Biotechnology Source Type: news
Cryopreservation of Red Blood Cells
Cryopreservation of red blood cell concentrates (RBCs) is an important method for maintaining an inventory of rare RBC units and managing special transfusion circumstances. The permeating additive glycerol is used as a cryoprotectant to protect RBCs against freezing damage. The use of thawed RBCs was hampered a 24-h outdating period due to potential bacterial contamination when a functionally open system was used for addition and removal of the glycerol. With the introduction of a functionally closed system for the glycerolization and deglycerolization of RBC units, extended post-thaw storage became possible. Here, we desc...
Source: Springer protocols feed by Biotechnology - December 2, 2014 Category: Biotechnology Source Type: news
