Determination of substitution positions in hyaluronic acid hydrogels using NMR and MS based methods
作者:Frida J. Wende、Suresh Gohil、Hotan Mojarradi、Thibaud Gerfaud、Lars I. Nord、Anders Karlsson、Jean-Guy Boiteau、Anne Helander Kenne、Corine Sandström
DOI:10.1016/j.carbpol.2015.09.112
日期:2016.1
the hydrogel. The substitution position of the cross-linker on the polysaccharide is another parameter that can influence hydrogel properties; hence methods for detailed structural analysis of the substitution pattern are required. NMR and LC-MS methods were developed to determine the positions and amounts of substitution of 1,4-butanediol diglycidyl ether (BDDE) on hyaluronic acid (HA), and for the
Method of determining nucleotide sequences in cells and of isolating nucleic acids from cells
申请人:NATIONAL RESEARCH DEVELOPMENT CORPORATION
公开号:EP0127327A1
公开(公告)日:1984-12-05
h has been a problem that nucleic acids present in cells have had to be isolated by difficult extraction procedures, using e.g. phenol or a protease, before they can be assayed.
The present invention provides an assay method which can be carried out on crude cell lysate, without the need for any such extraction.
The method comprises lysing the cells in a lysis buffer comprising a chaotropic agent for disrupting nucleoproteins and which inhibits the nuclease, e.g. a guanidinium salt present in the lysate in a concentration of at leat 3 M at the time of hybridisation, when DNA is being determined treating the lysate to single-strand it, contacting the lysate under hybridisation conditions with a polynucleotide having a nucleotide sequence complementary to a sequence present in the nucleic acid to be determined, and determining the extent of hybridisation, e.g. by a sandwich or competition assay.
Asimilar procedure can be used to extract the nucleic acid from the cell by a hybridisation method and isolate the desired nucleic acid from the resultant hybrid.
细胞中的核酸必须通过困难的提取程序(如使用苯酚或蛋白酶)分离出来,然后才能进行化验,这一直是个问题。
本发明提供了一种可在粗细胞裂解液上进行的检测方法,无需进行任何此类提取。
该方法包括在裂解缓冲液中裂解细胞,裂解缓冲液中含有一种可破坏核蛋白并抑制核酸酶的混沌剂,例如在测定 DNA 时,将裂解液处理成单链,在杂交条件下将裂解液与具有与待测核酸序列互补的核苷酸序列的多核苷酸接触,并测定杂交程度,例如通过夹心或竞争测定法。
可以使用类似的程序,通过杂交方法从细胞中提取核酸,并从杂交结果中分离出所需的核酸。
[EN] METHOD OF DETERMINING NUCLEOTIDE SEQUENCES IN CELLS AND OF ISOLATING NUCLEIC ACIDS FROM CELLS
申请人:——
公开号:WO1984004332A1
公开(公告)日:1984-11-08
(EN) It has been a problem that nucleic acids present in cells have had to be isolated by difficult extraction procedures, using e.g. phenol or a protease, before they can be assayed. The present invention provides an assay method which can be carried out on crude cell lysate, without the need for any such extraction. The method comprises lysing the cells in a lysis buffer comprising a chaotropic agent for disrupting nucleoproteins and which inhibits the nuclease, e.g. a guanidinium salt present in the lysate in a concentration of at least 3 M at the time of hybridisation, when DNA is being determined treating the lysate to single-strand it, contacting the lysate under hybridisation conditions with a polynucleotide having a nucleotide sequence complementary to a sequence present in the nucleic acid to be determined, and determining the extent of hybridisation, e.g. by a sandwich or competition assay. A similar procedure can be used to extract the nucleic acid from the cell by a hybridisation method and isolate the desired nucleic acid from the resultant hybrid. (FR) Avant de pouvoir analyser les acides nucléiques présents dans des cellules, le problème de leur isolation s"est toujours présenté à cause des procédés d"extraction difficiles ayant recours au phénol ou à une protéase par exemple. La présente invention fournit un procédé d"analyse qui peut être effectué sur un lysat de cellules brutes, sans qu"il soit nécessaire d"avoir recours à une telle extraction. Le procédé consiste à effectuer la lyse des cellules dans un tampon de lyse comprenant un agent chaotropique pour dissocier les nucléoprotéines et qui inhibe la nucléase, p. ex. un sel de guanidinium présent dans le lysat en une concentration d"au moins 3 M au moment de l"hybridation, lorsque l"ADN est en train d"être déterminé en traitant le lysat pour le rendre monobrin, en mettant en contact le lysat dans des conditions d"hybridation avec un polynucléotide ayant une séquence nucléotide complémentaire par rapport à une séquence présente dans l"acide nucléique à déterminer, et en déterminant le degré d"hybridation, p. ex. par une analyse en sandwich ou compétitive. Un procédé similaire peut être utilisé pour extraire l"acide nucléique de la cellule par un procédé d"hybridation et isoler l"acide nucléique désiré de l"hybride résultant.
Versatile and scalable synthesis of cyclic organic carbonates under organocatalytic continuous flow conditions
作者:Romaric Gérardy、Julien Estager、Patricia Luis、Damien P. Debecker、Jean-Christophe M. Monbaliu
DOI:10.1039/c9cy01659g
日期:——
epoxides. In this work, cyclic organic carbonates are prepared according to alternative sustainable and intensified continuousflow conditions from the corresponding 1,2-diols. The process utilizes dimethyl carbonate (DMC) as a low toxicity carbonation reagent and relies on the organocatalytic activity of widely available and cheap organic ammonium and phosphonium salts. Glycerol is selected as a model
The invention provides semi-solid systems for delivering biologically active materials that include a polymer comprising 1) one or more units of formula Ia, IIa, or IIIa:(formula Ia, IIa, IIIa) and 2) one or more units comprising polycaprolactone; wherein R and Ra have any of the values defined in the application.