Establishment a protocol for total RNA isolation from buffalo fresh and frozen semen for molecular applications

Andrologia ◽  
2020 ◽  
Vol 52 (4) ◽  
Author(s):  
Sally Ibrahim ◽  
Karima Gh. M. Mahmoud ◽  
Ahmed S. A. Sosa ◽  
Abdel aziz M. Sakr ◽  
Al‐shimaa Al‐H H. El‐Naby ◽  
...  
Keyword(s):  
Rna Isolation ◽  
Total Rna ◽  
Frozen Semen ◽  
Total Rna Isolation

Automated Solutions for Total RNA Isolation from Diverse Sample Types

2006 ◽  
Vol 11 (5) ◽  
pp. 309-313
Author(s):  
P SELLEY ◽  
J BRUNER ◽  
F KELLY ◽  
F MAURIO ◽  
M WATERS ◽  
...  
Keyword(s):  
Rna Isolation ◽  
Total Rna ◽  
Total Rna Isolation

New Data on Robustness of Gene Expression Signatures in Leukemia: Comparison of Three Distinct Total RNA Preparation Procedures.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4288-4288
Author(s):  
Marta Campo ◽  
Andrea Zangrando ◽  
Luca Trentin ◽  
Rui Li ◽  
Wei-min Liu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Expression Data ◽  
Rna Isolation ◽  
Expression Data ◽  
Microarray Technology ◽  
Isolation Method ◽  
Acute Leukemias ◽  
Preparation Methods ◽  
Total Rna ◽  
Total Rna Isolation

Abstract Gene expression microarrays had been used to classify known tumor types and various hematological malignancies (Yeoh et al, Cancer Cell 2002; Kohlmann et al, Genes Chromosomes Cancer 2003), enforcing the objective that microarray analysis could be introduced soon in the routine classification of cancer (Haferlach et al, Blood 2005). However, there’re still doubts about gene expression experiments performance in clinical laboratory diagnosis. For instance, the quality of starting material is a major concern in microarray technology and there are no data on the variation in gene expression profiles ensuing from different RNA extraction procedures. Here, as part of the internal multicenter MILE Study program, we assess the impact of different RNA preparation methods on gene expression data, analyzing 27 patients representative of nine different subtypes of pediatric acute leukemias. We compared the three currently most used protocols to isolate RNA for routine diagnosis (PCR assays) and microarray experiments. They are named as method A: lysis of mononuclear leukemia cells, followed by lysate homogeniziation, followed by total RNA isolation; method B: TRIzol RNA isolation, and method C: TRIzol RNA isolation followed by total RNA purification step. The methods were analyzed in triplicates for each sample (24) and additional three samples were performed in technical replicates of three data sets for each preparation (HG-U133 Plus 2.0). Method A results in better total RNA quality as demonstrated by 3′/5′ GAPD ratios and by RNA degradation plots. High comparability of gene expression data is found between samples in the same leukemia subclasses and collected with different RNA preparation methods thus demonstrating that sample preparation procedures do not impair the overall signal distribution. Unsupervised analyses showed clustering of samples first by each patient’s replicate conditions, then by leukemia type, and finally by leukemia lineage. In fact, B-ALL samples are clustered together, separately from T-ALL and AML, demonstrating that clustering reflects biological differences between leukemias and that the RNA isolation method is a secondary effect. Also, supervised cluster analyses highlight that samples are grouped depending on intra-lineage features (i.e. chromosomal aberrations) thus confirming the clustering organizations as reported in recent gene expression profiling studies of acute leukemias. Our study shows that biological features of pediatric acute leukemia classes largely exceed the variations between different total RNA sample preparation protocols. However, technical replicates analyses reveal that gene expression data from method A have the lowest degree of variation, are more reproducible and more precise as compared to the other two methods. Furthermore, compared to methods B and C, method A produces more differentially expressed probe sets between distinct leukemia classes and is therefore considered the more robust RNA isolation procedure for gene expression experiments using high-density microarray technology. We therefore conclude that method A (initial homogenization of the leukemia cell lysate followed by total RNA isolation) combined with a standardized microarray analysis protocol is highly reproducible and contributes to robustness of gene expression data and that this procedure is most practical for a routine laboratory use.

scholarly journals Optimization of total RNA isolation from Cochlodinium polykrikoides

2012 ◽  
Vol 25 (1) ◽  
pp. 59-64
Author(s):  
Kalnisha Devi Palani-Velu ◽  
Grace Joy Wei Lie Chin ◽  
Kenneth F. Rodrigues ◽  
Ann Anton
Keyword(s):  
Rna Isolation ◽  
Cochlodinium Polykrikoides ◽  
Total Rna ◽  
Total Rna Isolation

Total RNA isolation from stallion sperm and testis biopsies

2010 ◽  
Vol 74 (6) ◽  
pp. 1099-1106.e2 ◽  
Author(s):  
Pranab J. Das ◽  
Nandina Paria ◽  
Ashley Gustafson-Seabury ◽  
Monika Vishnoi ◽  
Sankar P. Chaki ◽  
...  
Keyword(s):  
Rna Isolation ◽  
Total Rna ◽  
Total Rna Isolation

Total RNA‐Isolation of Abdominal Hernia of Rats for Quantitative Real‐Time Reverse Transcription (RT) PCR Assays

2007 ◽  
Vol 38 (1) ◽  
pp. 87-93
Author(s):  
Christian Morsczeck ◽  
Michael Korenkov ◽  
Manfred Nagelschmidt ◽  
Domonkos Feher ◽  
Jörg Michael Schierholz
Keyword(s):  
Real Time ◽  
Reverse Transcription ◽  
Rna Isolation ◽  
Abdominal Hernia ◽  
Rt Pcr ◽  
Total Rna ◽  
Pcr Assays ◽  
Total Rna Isolation

Optimization of total RNA isolation from human urinary sediment

2016 ◽  
Vol 462 ◽  
pp. 158-161 ◽  
Author(s):  
Maria Beatriz Monteiro ◽  
Daniele Pereira Santos-Bezerra ◽  
Karina Thieme ◽  
Marisa Passarelli ◽  
Ubiratan Fabres Machado ◽  
...  
Keyword(s):  
Rna Isolation ◽  
Urinary Sediment ◽  
Total Rna ◽  
Total Rna Isolation

Total RNA Isolation Kit

2011 ◽  
Vol 2011 (3) ◽  
pp. pdb.kit54-pdb.kit54
Keyword(s):  
Rna Isolation ◽  
Total Rna ◽  
Total Rna Isolation

scholarly journals A simple and effective method for total RNA isolation of appressoria in Magnaporthe oryzae

2008 ◽  
Vol 9 (10) ◽  
pp. 811-817 ◽  
Author(s):  
Tong-bao Liu ◽  
Jian-ping Lu ◽  
Xiao-hong Liu ◽  
Hang Min ◽  
Fu-cheng Lin
Keyword(s):  
Magnaporthe Oryzae ◽  
Rna Isolation ◽  
Total Rna ◽  
Total Rna Isolation
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