OPTIMIZATION OF NUCLEIC ACID EXTRACTION AND RT-PCR FOR DETECTION OF GRAPEVINE FANLEAF VIRUS ISOLATES FROM VINEYARDS IN NORTH-WEST OF IRAN

Q: How can I download an article?

To download an article from SID, first log in to the site, search for the article title, and click on the 'Download Article' option.

Q: How can I download an ISI article?

To download an ISI article on SID, enter the keyword or article title in the search bar, view the relevant results, click on the desired article, and select the 'Download Article' option.

Q: How can I access the SID database?

To access the SID database, visit SID.ir, create an account, and log in to access scientific resources.

Q: Is downloading articles from SID free?

Some articles on SID are available for free, while others require payment. Details are specified on the article's page.

HAJIZADEH M.; SOKHANDAN BASHIR N.; NIKNAM GH.R.; NIKKHAH SH.

مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

Journal Paper

Paper Information

Journal: JOURNAL OF AGRICULTURAL KNOWLEDGE Year:2007 | Volume:16 | Issue:4 Page(s): 145-155

Download Full-Text

Persian Verion

View:

2,709

Download:

Cites:

Information Journal Paper

Title

OPTIMIZATION OF NUCLEIC ACID EXTRACTION AND RT-PCR FOR DETECTION OF GRAPEVINE FANLEAF VIRUS ISOLATES FROM VINEYARDS IN NORTH-WEST OF IRAN

Author(s)

HAJIZADEH M. | SOKHANDAN BASHIR N. | NIKNAM GH.R. | NIKKHAH SH. | Issue Writer Certificate

Keywords

CDNAQ3

GRAPEVINE FANLEA! VIRUSQ3

INHIBITORSQ3

PHENOL-CHLOROFORMQ3

POLYMERASE CHAIN REACTIONQ3

TOTAL RNAQ3

Abstract

One of the INHIBITORS of POLYMERASE CHAIN REACTION (PCR) in detecting fruit tree viruses including grapevine viruses is the presence of phenolic compounds and polysaccarides in the plant tissue. These INHIBITORS degrade the extracted nucleic acid and inactivate polymerases during PCR reaction. In this study, two protocols were examined to extract TOTAL RNA from the grapevines suspected to be infected by Grapevine fanleaf virus (GFLV). One protocol was based on PHENOL-CHLOROFORM extraction but the other did not use such solvents. In optimization of the RT-PCR reaction, synthesis of first strand CDNA was initially performed by either a virus-specific primer (S2515 or A3300) or oligo d(T)16 and subsequently PCR was undertaken with two primer sets CP433V/ CP912C or S2515/A33000-RT-PCR on PHENOL-CHLOROFORM extracted RNA did not result in any amplification from grapevine samples although the expected DNA fragments were amplified from OF LV- infected Chenopodium quinoa plants. Presumably, the phenol-based protocol was not able to remove the preventive substances from the extraction and they inhibited the RT-PCR reaction. However, RT with oligo d(T)16 on the RNA which was prepared without using phenol or 'chloroform from the grapevines and subsequent PCR with CP433V/ CP912C or S2515/A3300 resulted in amplification of the expected 480- or 810-bp DNA fragment, respectively. This study, for the first time, revealed the extent of OFLV distribution in vineyards of East Azarbaijan and Ardabil provinces in north-west of Iran plus providing ground for molecular characterization of the virus isolates.

Cites

No record.

References

SEROLOGICAL SUBGROUPING OF BASMENJ AND SHABESTAR ISOLATES OF CUCUMBER MOSAIC VIRUS AND APPLICATION OF RT-PCR FOR THE VIRUS DETECTION

Cite

APA: Copy

HAJIZADEH, M., SOKHANDAN BASHIR, N., NIKNAM, GH.R., & NIKKHAH, SH.. (2007). OPTIMIZATION OF NUCLEIC ACID EXTRACTION AND RT-PCR FOR DETECTION OF GRAPEVINE FANLEAF VIRUS ISOLATES FROM VINEYARDS IN NORTH-WEST OF IRAN. JOURNAL OF AGRICULTURAL KNOWLEDGE, 16(4), 145-155. SID. https://sid.ir/paper/28912/en

Vancouver: Copy

HAJIZADEH M., SOKHANDAN BASHIR N., NIKNAM GH.R., NIKKHAH SH.. OPTIMIZATION OF NUCLEIC ACID EXTRACTION AND RT-PCR FOR DETECTION OF GRAPEVINE FANLEAF VIRUS ISOLATES FROM VINEYARDS IN NORTH-WEST OF IRAN. JOURNAL OF AGRICULTURAL KNOWLEDGE[Internet]. 2007;16(4):145-155. Available from: https://sid.ir/paper/28912/en

IEEE: Copy

M. HAJIZADEH, N. SOKHANDAN BASHIR, GH.R. NIKNAM, and SH. NIKKHAH, “OPTIMIZATION OF NUCLEIC ACID EXTRACTION AND RT-PCR FOR DETECTION OF GRAPEVINE FANLEAF VIRUS ISOLATES FROM VINEYARDS IN NORTH-WEST OF IRAN,” JOURNAL OF AGRICULTURAL KNOWLEDGE, vol. 16, no. 4, pp. 145–155, 2007, [Online]. Available: https://sid.ir/paper/28912/en

Related Journal Papers