Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
1999-09-10
2001-09-18
Zitomer, Stephanie W. (Department: 1655)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C435S091500, C435S091510, C435S091100, C435S091200, C435S177000, C536S023100, C536S025400, C424S488000, C424S484000, C424S486000, C424S489000, C424S497000
Reexamination Certificate
active
06291179
ABSTRACT:
FIELD OF THE INVENTION
The present invention is directed to the storage, separation and analysis of genetic material. The invention is particularly suited for separating a genetic material from a sample containing multiple sources of a genetic material using a solid medium.
BACKGROUND OF THE INVENTION
The purification of genetic material from a crude sample for subsequent analysis can be laborious. Inadequate isolation of the genetic material from hemoglobin, proteins or other substance often associated with the generic material in a crude sample can inhibit or interfere with some analytical procedures such as, for example, polymerase chain reaction (“PCR”). Moreover, if the crude sample includes genetic material from multiple sources confusion of the genetic material analyzed can result.
Crude samples having multiple sources of genetic material include, for example, blood, feces, tissue fluids, plasmid containing bacteria, etc. The use of bacteria for propagation of plasmids is common in the study of genomics, analytic molecular biology, preparation molecular biology, etc. Methods for propagating plasmid containing bacteria are known. Common methods for storage of bacteria containing plasmids prior to analysis of the genetic material of the plasmid include, for example, filter paper, plastic, ceramic, semi-conductor, metal, etc.
Recently, new devices and methods for storage and purification of genetic material which are treated to protect the genetic material from degradation during storage and prior to analysis have become commercially available. Examples of such devices include products under the trade name FTA® available from Fitzco, Inc., Plymouth, Minn. Other examples are disclosed in U.S. Pat. Nos. 5,807,527; 5,756,126; and 5,496,562 and co-pending U.S. application Ser. No. 08/574,888. The disclosures of each of these patents and patent applications are incorporated herein by reference. Another example of a related product is IsoCode® available from Schleicher & Schuell, Keene, New Hampshire.
Purification and analysis of genetic material using a solid medium, such as those disclosed above, provide many advantages over wet storage methods. However, while these devices are suitable for storage and processing of genetic material, often times the genetic material stored in the form of a crude sample and may include genetic material derived from two or more different sources. For example, in the case of a bacterium containing a plasmid, genetic material from both the bacterium and plasmid are present. Thus, while a single sample may include genetic material from multiple sources, it is often advantageous to be able to distinguish the source of any particular piece of genetic material when the sample is analyzed.
Accordingly, there is a continuing need for products and methods which provide for purifying or separating a particular genetic material from a crude sample, including a sample containing genetic material from multiple sources.
SUMMARY OF THE INVENTION
The present application claims priority to U.S. Provisional Patent Application Ser. No. 60/099,715, filed Sep. 10, 1998, the entire disclosure of which is incorporated herein by reference.
The present invention is directed products and methods for purifying or separating a genetic material from a crude sample applied to a solid medium. In some embodiments, the invention provides for separating a selected genetic material from a sample containing genetic material from multiple sources.
It will be appreciated that throughout the specification, guidance may be provided through lists of examples. In each instance, the recited lists serve only as a representative group. It is not meant, however, that the lists are exclusive.
According to the invention, a sample of genetic material can be stored on a solid medium as described below. Once applied to the solid medium, non-genetic material typically associated with a crude sample, for example, proteins, fats, etc., can be removed from the dry solid medium using a cleaning solution. The dry solid medium can then be rinsed using a rinsing solution and the genetic sample analyzed in situ. Alternatively, the genetic material can be eluted from the solid medium and analyzed away from the solid medium.
In some embodiments, the invention further provides for purifying and removing a selected genetic material from a sample including genetic material from more than one source. According to this embodiment, the genetic material remaining on the solid medium can be further analyzed in situ, or, alternatively, the genetic material removed from the solid medium can further be analyzed away from the solid medium.
DETAILED DESCRIPTION OF THE INVENTION
The present invention is directed to products and methods which provide for separating genetic material derived from a crude sample that may contain genetic material from more than one source.
As used herein, the term “genetic material” includes a nucleotide sequence comprising two or more nucleotide bases including, for example, DNA, RNA, cDNA, oligonucleotides, entire genes, partial genes, entire genomes, etc. A “crude sample” refers to a sample of genetic material including components typically associated with the genetic material in vivo (e.g., proteins, fats, etc.) and includes, for example, a buccal swab, blood, serum, plasma, semen, feces, urine, cerebral spinal fluid, synovial fluid lymphatic fluid, etc. A “crude sample” also includes bacterial cultures, viral suspensions, etc. A “source” from which the genetic material is derived includes, for example, any type of eukaryotic or prokaryotic cell, viruses, plasmids, phages, archeobacteriae, plastids, cosmids, viroids, tissue fluids (blood, cerebrospinal fluid, lymphatic fluid, saliva, urine, etc.), etc. In one embodiment, the products and methods of the invention are particularly suited for purifying plasmid genetic material and separating genetic material of plasmids from bacteria containing the plasmids.
According to the invention, a sample containing genetic material can be applied to and stored on a medium comprising a support matrix such as paper (cellulose, nitrocellulose, etc.), plastic, ceramic, semi-conductor, metal, or other porous material or micro-indented surface or surface equipped with micropoles or microtrabeculae. In addition, the methods of the invention may also be used on genetic samples applied to and stored on a semi-solid matrix such as agar gel, agarose gel, polyacrylamide gel, etc.
The storage medium may also include one or more components which protect the stored genetic material from damage or degradation, denature proteins or which are useful for analysis of the stored genetic material. Examples of some such matrices are disclosed in U.S. Pat. Nos. 5,939,259; 5,807,527; 5,756,126; and 5,496,562; and co-pending U.S. Ser. No. 08/574,888.
As used herein, “analysis” of the genetic material includes methods used in the art for analyzing DNA or RNA such as sequencing, amplifying, hybridizing, probing, endonuclease restriction, ligation cloning, preparation for mass spectroscopy or irradiation, or any other procedure that is performed to obtain information about the genetic material. The genetic material can be analyzed immediately after taking the sample or stored for analysis at a later time.
According to the invention, some or all of the analysis to be performed on a particular source of genetic material can be analyzed in situ (i.e., on the medium) while source(s) not to be analyzed are removed from the medium prior to analysis. Alternatively, the genetic material to be analyzed can be removed from the medium, for example, the sources of genetic material which are not to be analyzed can remain on the medium and the genetic material to be analyzed removed from the medium prior to analysis. Thus, to analyze a particular source of genetic material from a multiple source sample, the methods herein described provide for selective removal from the solid medium of either the genetic material to be analyzed or the genetic material (and/or associated non-genetic material)
Kohn & Associates
Whatman PLC
Wilder Cynthia B.
Zitomer Stephanie W.
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