摘要:
The present application is directed to a method for performing a bisulfite reaction to determine methylation positions in a nucleic acid, i.e. methylated and non-methylated cytosines, whereby the nucleic acid is incubated in a solution comprising the nucleic acid for a time period of 1.5 to 3.5 hours at a temperature between 70 and 90° C., whereby the concentration of bisulfite in the solution is between 3 M and 6.25 M and whereby the pH value of the solution is between 5.0 and 6.0 whereby the nucleic acid, i.e. the cytosine bases in the nucleic acid, are deaminated. Then the solution comprising the deaminated nucleic acid is desulfonated and preferably desalted. The application is further related to a solution comprising bisulfite with a certain pH and uses thereof as well as a kit comprising the solution.
摘要:
The present application is directed to a method for performing a bisulfite reaction to determine methylation positions in a nucleic acid, i.e. methylated and non-methylated cytosines, whereby the nucleic acid is bound to a solid phase during the deamination and/or desulfonation step of the bisulfite reaction. The solid phase is preferably a material comprising glass or silica, more preferably a glass fleece, glass membrane or a magnetic glass particle. Further, the use of a solid phase for binding a nucleic acid during the deamination and/or desulfonation step of the bisulfite reaction is disclosed and a kit containing a bisulfite reagent and a solid phase.
摘要:
The invention is related to a method for amplifying a methylated target nucleic acid in a sample while avoiding amplification of a non-methylated target nucleic acid by inactivating it. This is accomplished by a restriction enzyme digest after bisulfite treatment of the target nucleic acid. The invention is further related to the use of a restriction enzyme to avoid amplification of a non-methylated target nucleic acid while amplifying a methylated target nucleic acid in a sample and kits for performing the methods according to the invention.
摘要:
The invention is related to a method for amplifying a methylated target nucleic acid in a sample while avoiding amplification of a non-methylated target nucleic acid by inactivating it. This is accomplished by a restriction enzyme digest after bisulfite treatment of the target nucleic acid. The invention is further related to the use of a restriction enzyme to avoid amplification of a non-methylated target nucleic acid while amplifying a methylated target nucleic acid in a sample and kits for performing the methods according to the invention.
摘要:
The invention concerns a method for determining antigen-specific antibodies of a particular immunoglobulin class in a sample by means of an immunoassay in an array format in which various binding partners Bnx are bound on different discrete areas on a support where Bnx in each case contain the various antigens that are able to specifically bind to the antibodies to be detected, by incubating the support with the sample and a binding partner B2 which carries a label and subsequently detecting the label on the respective discrete areas wherein B2 specifically binds antibodies of a certain immunoglobulin class that have been bound in an antigen-specific manner.
摘要:
Methods for determining an analyte in a sample by immunoassay in a one-step format without performing washing steps are described. The method includes a first analyte-specific receptor that contains at least two binding sites for the analyte, and a second analyte-specific receptor that selectively binds to an aggregate arrangement of at least two analyte molecules bound to the first receptor.
摘要:
The invention relates to a method for determining an analyte in a sample by means of an assay that can be carried out in a one-step format without performing washing steps. The method includes a first analyte-specific receptor that contains at least one bonding point for the analyte, e.g., several antibodies on a particle, as well as a second analyte-specific receptor that can selectively bond to an arrangement comprising at least two analyte molecules which are bound to the first receptor. An antibody sandwich immune assay is described in which the detection is based on electrochemiluminescence.
摘要:
The invention is related to the detection of a methylated cytosine in a nucleic acid wherein guanidinium hydrogen sulfite is used for the preparation of a solution containing guanidinium ions and sulfite ions and subsequent modification of the nucleic acid. Thereby, a non-methylated cytosine is converted to uracil. The invention further discloses kits for performing the methods of the invention.
摘要:
The present application is directed to a method for performing a bisulfite reaction to determine methylation positions in a nucleic acid, i.e. methylated and non-methylated cytosines. In this method, the nucleic acid is bisulfite treated and is bound to a solid phase when an alkaline solution is added for desulfonation and elution of the nucleic acid from the solid phase. The nucleic acid can be amplified in an additional step. The solid phase is preferably a material comprising glass or silica, more preferably a glass fleece, glass membrane or a magnetic glass particle. Further, several uses of the alkaline solution for bisulfite treatment are disclosed and a kit containing a bisulfite reagent, a solid phase and an alkaline solution.
摘要:
The invention concerns a process for preparing biological samples for the subsequent detection of an analyte. In particular, the invention relates to a process for the isolation of a nucleic acid in a sample using a suspension of magnetic glass particles. In addition, kits and apparatuses containing magnetic glass particles for sample preparation are provided.