Abstract：
:Construction of a random ssDNA sublibrary is an important step of the aptamer screening process. The available construction methods include asymmetric PCR, biotin-streptavidin separation, and lambda exonuclease digestions, in which PCR amplification is a key step. The main drawback of PCR amplification is overamplification increasing nonspecific hybridization among different products and by-products, which may cause the loss of potential high-quality aptamers, inefficient screening, and even screening failure. Cycle number optimization in PCR amplification is the main way to avoid overamplification but does not fundamentally eliminate the nonspecific hybridization, and the decreased cycle number may lead to insufficient product amounts. Here, we developed a new method, "asymmetric emulsion PCR," which could overcome the shortcomings of conventional PCR. In asymmetric emulsion PCR, different templates were separated by emulsion particles, allowing single-molecule PCR, in which each template was separately amplified, and the nonspecific hybridization was avoided. Overamplification or formation of by-products was not observed. The method is so simple that direct amplification of 40 or more cycles can provide a high-quality ssDNA library. Therefore, the asymmetric emulsion PCR would improve the screening efficiency of systematic evolution of ligands by exponential enrichment.
journal_name
Biotechnol Appl Biochemjournal_title
Biotechnology and applied biochemistryauthors
Shao K,Shi X,Zhu X,Cui L,Shao Q,Ma Ddoi
10.1002/bab.1467subject
Has Abstractpub_date
2017-03-01 00:00:00pages
239-243issue
2eissn
0885-4513issn
1470-8744journal_volume
64pub_type
杂志文章abstract：:Various approaches for removal of high-abundance components in body fluids are currently available. While most methods are constructed for plasma depletion, there is a need for body-fluid-specific strategies. The aim of the present study was to design an affinity matrix suitable for the depletion of high-abundance pro...
journal_title：Biotechnology and applied biochemistry
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doi：10.1042/BA20080015
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abstract：:Two monoclonal antibodies against human myeloperoxidase, designated 3-2H3 and 4-2C11, were produced and characterized. Both bound to the native enzyme, but neither bound to the denatured enzyme or to its two denatured subunits. 4-2C11 bound to the three types of leukocyte myeloperoxidase, I, II, and III, as well as to...
journal_title：Biotechnology and applied biochemistry
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更新日期：1986-12-01 00:00:00
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journal_title：Biotechnology and applied biochemistry
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doi：10.1042/ba19990102
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abstract：:Bovine serum albumin (BSA) microspheres with an average diameter of 12.5 micron were prepared by crosslinking of BSA molecules with glutaraldehyde in the presence of polymethylmethacrylate dissolved in chloroform-toluene. Trypsin and anti-human IgG antibody were immobilized onto their surfaces by the glutaraldehyde-ac...
journal_title：Biotechnology and applied biochemistry
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更新日期：1988-02-01 00:00:00
abstract：:The aspartic proteinase gene of Mucor pusillus rennin expressed in Pichia pastoris was characterized in terms of structural and conformational stability induced by temperature. This enzyme is 12% glycosylated, with a similar specific activity to the native fungal enzyme. The secondary structure determined by CD is mai...
journal_title：Biotechnology and applied biochemistry
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doi：10.1042/ba19990094
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journal_title：Biotechnology and applied biochemistry
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journal_title：Biotechnology and applied biochemistry
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journal_title：Biotechnology and applied biochemistry
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journal_title：Biotechnology and applied biochemistry
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journal_title：Biotechnology and applied biochemistry
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更新日期：2013-01-01 00:00:00
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pub_type： 杂志文章
doi：10.1002/bab.1981
更新日期：2020-07-04 00:00:00
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journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1002/bab.1386
更新日期：2016-05-01 00:00:00
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journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：10.1042/BA20050114
更新日期：2005-12-01 00:00:00
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journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：
更新日期：1991-04-01 00:00:00
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journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：
更新日期：1988-06-01 00:00:00
abstract：:High yields of extracellular alpha-galactosidase from fungal cultures were obtained by inducing enzyme production with guar gum (a galactomannan obtained from the seeds of Cyamopsis tetragonobola) as the sole carbon source. An alpha-galactosidase was isolated from the culture medium of Penicillium ochrochloron culture...
journal_title：Biotechnology and applied biochemistry
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doi：
更新日期：1993-06-01 00:00:00
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journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章,评审
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更新日期：2008-01-01 00:00:00
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journal_title：Biotechnology and applied biochemistry
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doi：
更新日期：1995-10-01 00:00:00
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journal_title：Biotechnology and applied biochemistry
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更新日期：2006-05-01 00:00:00
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journal_title：Biotechnology and applied biochemistry
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更新日期：2002-06-01 00:00:00
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journal_title：Biotechnology and applied biochemistry
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更新日期：2010-02-11 00:00:00
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journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
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更新日期：2015-11-01 00:00:00
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journal_title：Biotechnology and applied biochemistry
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更新日期：2000-04-01 00:00:00
abstract：:Glutaraldehyde crosslinked, immunoprecipitated gulonolactone oxidase, injected intraperitoneally, has significant catalytic activity and is capable of providing long-term therapeutic benefit for the enzyme deficiency disease scurvy. The enzyme is tolerated even in repetitive doses. In the present study, however, we ha...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：
更新日期：1986-04-01 00:00:00
abstract：:As a model of cell-cell recognition processes, the association processes of a polysaccharide (mannan)-carrying liposome with a lectin (Concanavalin A, Con A)-carrying polymerized liposome were followed by turbidimetry. The association process was strongly inhibited by the addition of a low molecular weight sugar, meth...
journal_title：Biotechnology and applied biochemistry
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doi：10.1111/j.1470-8744.1992.tb00195.x
更新日期：1992-02-01 00:00:00
abstract：:We have analysed the status and transcriptional activity of the bovine papillomavirus-I (complete BPV-I genome)-based expression vector pCES in CI27i-cell-line-derived 3TI cells used for the industrial production of recombinant human erythropoietin (rhuEpo). Complete tandem head-to-tail integration of about 600 vector...
journal_title：Biotechnology and applied biochemistry
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doi：
更新日期：1996-02-01 00:00:00
abstract：:The use of glutaraldehyde (GTA) for cross-linking biological tissue implants has a number of undesirable side effects, including cytotoxicity and induction of calcification. In an attempt to find an improved cross-linking agent for tissue implants, we have evaluated a number of cross-linking procedures shown previousl...
journal_title：Biotechnology and applied biochemistry
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更新日期：1993-02-01 00:00:00
abstract：:The results from molecular mass determinations and amino acid analyses for prostate-specific antigen (PSA), alpha 1-antichymotrypsin (ACT) and PSA-ACT complexed in vitro are reported. Molecular masses for two separate PSA-ACT lots were determined by matrix-assisted laser desorption ionization mass spectroscopy (MALDI-...
journal_title：Biotechnology and applied biochemistry
pub_type： 杂志文章
doi：
更新日期：1998-06-01 00:00:00
