Analysis of RNA by Northern and Slot Blot Hybridization

1997 ◽  
Vol 37 (1) ◽  
Author(s):  
Terry Brown ◽  
Karol Mackey
Keyword(s):  
Blot Hybridization ◽  
Slot Blot ◽  
Slot Blot Hybridization

Using genomic slot blot hybridization to assess intergeneric Saccharum x Erianthus hybrids (Andropogoneae – Saccharinae)

Genome ◽  
1997 ◽  
Vol 40 (4) ◽  
pp. 428-432 ◽  
Author(s):  
P. Besse ◽  
C. L. McIntyre ◽  
D. M. Burner ◽  
C. G. de Almeida
Keyword(s):  
Genomic Dna ◽  
Blot Hybridization ◽  
Female Parent ◽  
Saccharum Officinarum ◽  
Rapid Screening ◽  
Hybridization Technique ◽  
Slot Blot ◽  
Saccharum Species ◽  
Slot Blot Hybridization

The use of genomic slot blot hybridization enabled the differentiation of hybrids from selfs in Saccharum × Erianthus intergeneric crosses in which Saccharum was used as the female parent. Based on the genomic in situ hybridization technique, slot blots of DNA from the parents and the progeny were blocked with the Saccharum parent DNA and hybridized with the labelled male Erianthus genomic DNA. This technique allowed a rapid screening for hybrids and was sensitive enough to detect a 1/20 dilution of Erianthus in Saccharum DNA, which should enable the detection of most partial hybrids. The genomic slot blot hybridization technique was shown to be potentially useful for assessing crosses involving Saccharum species with either Old World Erianthus section Ripidium or North American Erianthus (= Saccharum) species. The effectiveness of the technique was assessed on 144 progeny of a Saccharum officinarum × Erianthus arundinaceus cross, revealing that 43% of the progeny were selfs. The importance of this test as a tool to support intergeneric breeding programs is discussed.Key words: slot blot, Erianthus, genomic DNA, Saccharum, sugarcane.

scholarly journals Using a CCD Camera Imaging System as a Recording Device to Quantify Human DNA by Slot Blot Hybridization

BioTechniques ◽  
2001 ◽  
Vol 30 (3) ◽  
pp. 680-685 ◽  
Author(s):  
Bruce Budowle ◽  
William R. Hudlow ◽  
Steven B. Lee ◽  
Leonard Klevan
Keyword(s):  
Imaging System ◽  
Blot Hybridization ◽  
Ccd Camera ◽  
Recording Device ◽  
Slot Blot ◽  
Camera Imaging ◽  
Human Dna ◽  
Slot Blot Hybridization

Sulfated glycoprotein-1 (SGP-1) expression in ovine endometrium during the oestrous cycle and early pregnancy

1995 ◽  
Vol 7 (5) ◽  
pp. 1053 ◽  
Author(s):  
TE Spencer ◽  
GH Graf ◽  
FW Bazer
Keyword(s):  
Early Pregnancy ◽  
Blot Hybridization ◽  
Immunohistochemical Localization ◽  
Oestrous Cycle ◽  
Northern Hybridization ◽  
Mrna Levels ◽  
Slot Blot ◽  
Slot Blot Hybridization ◽  
Endometrial Epithelium ◽  
Sulfated Glycoprotein

This study determined effects of day of oestrous cycle and early pregnancy on sulfated glycoprotein-1 (SGP-1) expression in ovine endometrium. A 364-bp clone of the ovine SGP-1 mRNA was amplified from reverse transcribed Day-15 cyclic endometrial mRNA using the polymerase chain reaction (PCR) and primers specific for the rat SGP-1 mRNA sequence. Nucleotide sequence of the ovine SGP-1 cDNA shared significant identity with rat SGP-1 and human prosaposin. Ewes (n = 40) were hysterectomized on either Day 1, 6, 11, 13 or 15 of the oestrous cycle or on Day 11, 13, 15, 17 or 25 of early pregnancy. Total cellular RNA was isolated from endometrium and subjected to Northern and slot blot hybridization analyses using an antisense cRNA probe transcribed from the ovine SGP-1 cDNA clone. A single 2.6-kb mRNA transcript was detected by Northern hybridization analyses. Slot blot hybridization analyses indicated that steady-state levels of endometrial SGP-1 mRNA varied during the oestrous cycle (cubic, P < 0.02) and increased between Day 11 and Day 25 of early pregnancy (linear, P < 0.01). On Days 11, 13 and 15, endometrial SGP-1 mRNA levels were greater in pregnant ewes than in cyclic ewes (day x pregnancy status, P < 0.01). Immunohistochemical localization of SGP-1 in uterine tissues with rabbit anti-rat SGP-1 antibody revealed intense immunoreactivity associated primarily with the endometrial epithelium. These results indicate that the ovine endometrium expresses SGP-1, a prosaposin, and that SGP-1 expression varies during the oestrous cycle and is enhanced by the conceptus. The presence of SGP-1 in the endometrium suggests intracellular and extracellular roles for this protein in glycosphingolipid metabolism or transport in the uterine environment.

Identification of rare Candida species and other yeasts by polymerase chain reaction and slot blot hybridization

2000 ◽  
Vol 38 (4) ◽  
pp. 207-212 ◽  
Author(s):  
Juergen Loeffler ◽  
Holger Hebart ◽  
Stella Magga ◽  
Diethard Schmidt ◽  
Lena Klingspor ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Candida Species ◽  
Blot Hybridization ◽  
Slot Blot ◽  
Chain Reaction ◽  
Slot Blot Hybridization ◽  
Polymerase Chain

scholarly journals Early occurrence of human cytomegalovirus infection after bone marrow transplantation as demonstrated by the polymerase chain reaction technique

Blood ◽  
1991 ◽  
Vol 77 (5) ◽  
pp. 1104-1110 ◽  
Author(s):  
H Einsele ◽  
M Steidle ◽  
A Vallbracht ◽  
JG Saal ◽  
G Ehninger ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Polymerase Chain Reaction ◽  
Bone Marrow Transplantation ◽  
Blot Hybridization ◽  
Slot Blot ◽  
Chain Reaction ◽  
Biopsy Specimens ◽  
Slot Blot Hybridization ◽  
Polymerase Chain ◽  
Pcr Techniques

Abstract Twenty-eight patients undergoing bone marrow transplantation (BMT) were followed-up at weekly intervals from day -10 to discharge from hospital after BMT for human cytomegalovirus (HCMV) infection using polymerase chain reaction (PCR), slot-blot hybridization, and conventional virus culture. High specificity of the PCR assay applied could be shown by failure to amplify DNA extracted from a wide range of other viruses frequently infecting marrow transplant recipients. The PCR technique allowed us to diagnose viremia and viruria in 20 (83%) of 24 seropositive patients after BMT, whereas culture assays showed 16 (67%) of 24 of these patients to be viruric and 9 (37%) of 24 cases to be viremic. Slot-blot hybridization showed a frequency of viruria and viremia in 12 (50%) of 24 seropositive patients. By application of PCR techniques, HCMV detection could be achieved even in the very early posttransplant period. HCMV was detected in five patients even before the onset of clinical symptoms of acute graft-versus-host disease. Analysis by PCR techniques of 33 organ biopsy specimens from patients after BMT showed the presence of HCMV in 13 of 14 liver samples obtained from patients with HCMV viremia; three liver specimens from patients without viremia were negative by all the techniques applied. HCMV could also be demonstrated in postmortem lung biopsy specimens from all patients (n = 10) with interstitial pneumonia.

scholarly journals Chemiluminescent Detection of Infectious Bursal Disease Virus with a PCR-Generated Nonradiolabeled Probe

1993 ◽  
Vol 5 (2) ◽  
pp. 166-173 ◽  
Author(s):  
A. Akin ◽  
C. C. Wu ◽  
T. L. Lin ◽  
R. W. Keirs
Keyword(s):  
Disease Virus ◽  
Infectious Bursal Disease Virus ◽  
Salmonella Enteritidis ◽  
Blot Hybridization ◽  
Infectious Bursal Disease ◽  
Slot Blot ◽  
Slot Blot Hybridization ◽  
Bursal Disease ◽  
Serotype 1 ◽  
Labeled Probe

A polymerase chain reaction (PCR)-generated digoxigenin-labeled nonradioactive oligonucleotide probe was developed and utilized in slot-blot hybridization coupled with chemiluminescence for the detection of infectious bursal disease virus (IBDV). The probe was prepared from the RNA of the standard challenge strain (STC) of IBDV serotype 1 by reverse transcription followed by 2 PCR amplifications with 2 separate sets of primers. RNA of STC viruses prepared from bursae infected with STC viruses was subjected to the first PCR with the outer primers V8 and V9 that amplified a 607-base pair (bp) segment. The PCR product from the first PCR was eluted following agarose gel electrophoresis and subjected to the second PCR with the nested primers V6 and V7 that flanked a 351-bp segment. In the second PCR, dTTP was substituted by digoxigenin-11-dUTP in the PCR reaction mixture so that the amplified 351-bp DNA products were labeled with digoxigenin. The specificity of the PCR-generated digoxigenin-labeled probe was tested on different strains of IBDV, several unrelated avian viruses, and bacteria by slot-blot hybridization assay. Hybridization was detected by chemiluminescence. The sensitivity of the probe was assayed using lo-fold serial dilutions of purified RNA from the STC strain of IBDV. The PCR-generated digoxigenin-labeled probe hybridized with genomic RNA of STC and variant strains A, D, E, G, and GLS-5 of IBDV serotype 1 but not OH strain of IBDV serotype 2. The probe did not react with avian reovirus, infectious bronchitis virus, Salmonella enteritidis, Escherichia coli, or Staphylococcus aureus. The probe was very sensitive, and as little as 72 fg of RNA from the STC strain of IBDV could be detected. The results indicate that this PCR-generated digoxigenin-labeled nonisotopic probe is specific for IBDV and may be utilized in a diagnostic assay for all IBDV serotype 1 strains.

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