his.sePublikationer
Ändra sökning
RefereraExporteraLänk till posten
Permanent länk

Direktlänk
Referera
Referensformat
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Annat format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annat språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf
Evaluation of different modifications of TRIzol Reagent method for total RNA isolation from bull spermatozoa
Högskolan i Skövde, Institutionen för vård och natur. (The Systems Biology Research Center)ORCID-id: 0000-0002-0785-8945
2013 (Engelska)Självständigt arbete på avancerad nivå (magisterexamen), 20 poäng / 30 hpStudentuppsats (Examensarbete)
Abstract [en]

Transcriptional profile of spermatozoa can be used to assess male fertility; differences in spermatozoa attributes require adjustment of protocols for RNA isolation. Ten RNA precipitation modifications of heated TRIzol protocol were applied to total RNA extraction form bull spermatozoa. Motile spermatozoa were isolated from cryopreserved semen straws prepared from an ejaculate of a fertile bull. To evaluate total RNA purity and quantity, Ribogreen RNA detection system and Nanodrop 2000 were used. In RiboGreen assay, a consistent standard curve was not obtained, thus the measurements were ignored. Total RNA purity and concentrations also were assessed using Nanodrop. The A260/280 ratio obtained from Nanodrop showed a good range in the isolated total RNA samples (1.9±0.4). However, all RNA samples presented very low A260/230 ratios (≥1.26). It might be the effect of low concentration of the isolated total RNA. Statistical analysis of total RNA concentrations showed using manufacturer standard protocol had the highest concentration yield. Using either ethanol or Isopropanol as RNA precipitant and changing the incubation temperature had not significant effect on RNA concentration yields. However incubating samples in freezer overnight lowered the total RNA concentration. Using glycogen as a RNA carrier increased the total RNA precipitation yield. In other similar studies the highest total RNA concentration was obtained by using RNA carrier. Validating these findings by qRT-PCR and Bioanalyser might be helpful to enhance the total RNA extraction yield from bull sperm as well as for subsequent genomic studies of bull fertility.

Ort, förlag, år, upplaga, sidor
2013. , s. 18
Nyckelord [en]
total RNA precipitation, spermatozoa, RNA extra ction, bull fertility, TRIzol reagent
Nationell ämneskategori
Biomedicinsk laboratorievetenskap/teknologi
Identifikatorer
URN: urn:nbn:se:his:diva-8283OAI: oai:DiVA.org:his-8283DiVA, id: diva2:631403
Ämne / kurs
Biomedicin/medicinsk vetenskap
Utbildningsprogram
Biomedicin - magisterprogram
Presentation
2013-05-31, G211, University of Skövde, Skövde, 13:30 (Engelska)
Uppsök
bio-/geovetenskap
Handledare
Examinatorer
Projekt
Correlation between bull fertility and miRNA signature of the sperm – Prediction of Bull FertilityTillgänglig från: 2013-11-19 Skapad: 2013-06-20 Senast uppdaterad: 2013-11-19Bibliografiskt granskad

Open Access i DiVA

Fulltext saknas i DiVA

Sök vidare i DiVA

Av författaren/redaktören
Etezadi Amoli, Mahmood Reza
Av organisationen
Institutionen för vård och natur
Biomedicinsk laboratorievetenskap/teknologi

Sök vidare utanför DiVA

GoogleGoogle Scholar

urn-nbn

Altmetricpoäng

urn-nbn
Totalt: 367 träffar
RefereraExporteraLänk till posten
Permanent länk

Direktlänk
Referera
Referensformat
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Annat format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annat språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf