Susan Rigby - Acton MA, US Heather P. O'Keefe - Lexington MA, US Henrik Stender - Gentofte, DK
Assignee:
Boston Probes, Inc. - Bedford MA
International Classification:
C12Q001/68 C12P019/34 C07H021/02 C07H021/04
US Classification:
435 6, 435 912, 536 235, 536 2431, 536 2433
Abstract:
This invention is directed to a rapid and simple method for determining organisms and/or cells in a sample. Generally, the method is directed to the use of molecular probes to selectively stain the organisms and/or cells for determination wherein growth medium, fixative reagents and/or excess molecular probes need not be separated before a determination is made.
Methods For The Detection, Identification, And/Or Enumeration Of Yeast, Particularly In Wine
This invention is related to novel probes, probe sets, methods and kits pertaining to the detection, identification and/or quantitation of yeasts and particularly (a. k. a. ) an organism that spoils wine. Preferred probes for the detection of one or more species of the genus comprise a probing nucleobase sequence, at least a portion of which is selected from the group consisting of: AGC-GGG-TCT-ATT-AGA (Seq. ID No. 1); CCA-GGT-GAG-GGT-CGC (Seq. ID No. 2); CGG-TTG-CCC-GAT-TTC (Seq. ID No. 3); TCG-CCT-TCC-TCC-TCT (Seq. ID No. 4); CGG-TCT-CCA-GCG-ATT (Seq. ID No. 5); CAC-AAG-ATG-TCC-GCG (Seq. ID No. 6); GCG-GGC-ACT-AAT-TGA (Seq. ID No. 7); CAT-CCA-CGA-GGA-ACG (Seq. ID No. 8); GTG-TAA-ACC-AGG-TGC (Seq. ID No. 9); ATG-GCT-CCC-AGA-ACC (Seq. ID No. 10) and GAC-AGA-ATC-GAA-GGG (Seq. ID No. 11).
Methods For The Analysis Of Microcolonies Of Bacteria And Yeast
This invention is related to methods pertaining to the determination of bacteria and yeast wherein the methods comprise: a) filtering a sample containing bacteria and yeast through at least one filter; b) treating the filter or filters with a medium which facilitates the growth of the bacteria and yeast; c) growing the bacteria and yeast on the filter or filters to thereby produce colonies or microcolonies of the bacteria and yeast; d) fixing the colonies or microcolonies to the filter or filters; e) treating each filter under suitable hybridization conditions with one or more enzyme labeled PNA probes wherein, each PNA probe comprises a probing nucleobase sequence complementary to a target sequence in the nucleic acid of the bacteria and/or yeast; f) removing excess enzyme labeled PNA probe from each of the filters; and g) determining enzyme activity remaining on each filter to thereby identify or enumerate colonies or microcolonies on the filter or filters.
Methods And Kit For Hybridization Analysis Using Peptide Nucleic Acid Probes
MARTIN FUCHS - UXBRIDGE MA, US MICHAEL EGHOLM - LEXINGTON MA, US HEATHER O'KEEFE - LEXINGTON MA, US
International Classification:
C12Q001/00 G01N031/00 G01N033/53 C25D021/12
US Classification:
435/004000, 436/516000, 435/007100, 205/081000
Abstract:
A method composition and apparatus for the hybridization and separation of molecules having a desired target sequence in a sample by contacting a sample of single stranded nucleic acids with a detectable PNA probe having a sequence complementary to the target sequence so that the target sequence, if present, will hybridize with the detectable probe to form a detectable duplex, and then separating the duplex in a denaturing medium from unbound sample components by electrophoresis. The invention also relates to methods compositions and apparatus for the hybridization and separation of molecules having a desired target sequence in a mixed sample of single stranded nucleic acids and their complementary strands by contacting the sample with a detectable PNA probe.
Methods And Kit For Hybridization Anaylsis Using Peptide Nucleic Acid Probes
Martin Fuchs - Uxbridge MA, US Michael Egholm - Woodbridge CT, US Heather O'Keefe - Lexington MA, US
International Classification:
C12Q001/68 C07K014/47
US Classification:
435006000, 530350000
Abstract:
A method composition and apparatus for the hybridization and separation of molecules having a desired target sequence in a sample by contacting a sample of single stranded nucleic acids with a detectable PNA probe having a sequence complementary to the target sequence so that the target sequence, if present, will hybridize with the detectable probe to form a detectable duplex, and then separating the duplex in a denaturing medium from unbound sample components by electrophoresis. The invention also relates to methods compositions and apparatus for the hybridization and separation of molecules having a desired target sequence in a mixed sample of single stranded nucleic acids and their complementary strands by contacting the sample with a detectable PNA probe.
Pna Probes, Probe Sets, Methods And Kits Pertaining To The Universal Detection Of Bacteria And Eucarya
This invention is related to novel PNA probes, probe sets, methods and kits pertaining to the universal detection of bacteria and/or eucarya. Preferred universal probes for the detection of bacteria comprise a probing nucleobase sequence selected from the group consisting of CTG-CCT-CCC-GTA-GGA; TAC-CAG-GGT-ATC-TAA-T; CAC-GAG-CTG-ACG-ACA and CCG-ACA-AGG-AAT-TTC. Preferred universal probes for the detection of eucarya comprise a probing nucleobase sequence selected from the group consisting of ACC-AGA-CTT-GCC-CTC-C; GGG-CAT-CAC-AGA-CCT-G; TAG-AAA-GGG-CAG-GGA and TAC-AAA-GGG-CAG-GGA. The PNA probes, probe sets, methods and kits of this invention are particularly well suited for use in multiplex PNA-FISH assays wherein the bacteria and/or eucarya in a sample can be individually detected, identified or quantitated. Using exemplary assays described herein, the total number of colony forming units (CFU) of bacteria and/or eucarya can be rapidly determined.