LIPOFECTAMINE DOES NOT ENHANCE BOVINE SPERM TRANSFECTION EFFICIENCY

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EGHBAL SAEID SH.; GHAEDI K.; NASRESFAHANI MOHAMMAD HOSSEIN; AMIR MOZAFARI N.

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Journal Paper

Paper Information

Journal: Cell Journal (Yakhteh) Year:2008 | Volume:10 | Issue:SUPPLEMENT 1 Page(s): 48-48

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Title

LIPOFECTAMINE DOES NOT ENHANCE BOVINE SPERM TRANSFECTION EFFICIENCY

Author(s)

EGHBAL SAEID SH. | GHAEDI K. | NASRESFAHANI MOHAMMAD HOSSEIN | AMIR MOZAFARI N.

Keywords

TRANSFECTION

LIPOFECTAMINE

DMSO

SPERM AND BOVINE

Abstract

Objective: (i) if using chemical reagents such as DMSO and LIPOFECTAMINE™2000 increase bovine sperm TRANSFECTION, (ii) if motile spermatozoa have a greater TRANSFECTION rate than immotile ones, and (iii) if increasing sperm- DNA incubation time enhance sperm TRANSFECTION rate.Materials and Methods: Motile bull sperms were allowed to swim up at 38oC for 90 min. The rhodamine labeled pGeneGrip-Lac-Z vector (Genelantic, USA) was used for direct detection of transfected cells under epiflourescence microscope. One and three microlitersof LIPOFECTAMINE2000 were added separately into two tubes containing 20μl of SP-TALP medium (BSA and antibiotic free) and stored at room temperature for 30 min. One microgram of the vector was added to liposomes and stored at room temperature for a further 90 min to perform 1:1 and 1:3 (DNA: LIPOFECTAMINE) ratios. one mg of naked DNA was added into SP-TALP medium as control group and supplemented with 1% and 3% Dimethyl sulfoxide(DMSO) as DMSO groups. for all groups, 1×106 fully capacitated spermatozoa was added to each tube and the total volume adjusted to 30ml with SP-TALP medium (containing 6% BSA) and incubated at 38oC for 60 min. Sperm-DNA mixtures were divided into two groups, treated with or without DNaseI (5 UI) for 30 min and used for fluorescence detection. In another

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APA: Copy

EGHBAL SAEID, SH., GHAEDI, K., NASRESFAHANI, MOHAMMAD HOSSEIN, & AMIR MOZAFARI, N.. (2008). LIPOFECTAMINE DOES NOT ENHANCE BOVINE SPERM TRANSFECTION EFFICIENCY. CELL JOURNAL (YAKHTEH), 10(SUPPLEMENT 1), 48-48. SID. https://sid.ir/paper/542430/en

Vancouver: Copy

EGHBAL SAEID SH., GHAEDI K., NASRESFAHANI MOHAMMAD HOSSEIN, AMIR MOZAFARI N.. LIPOFECTAMINE DOES NOT ENHANCE BOVINE SPERM TRANSFECTION EFFICIENCY. CELL JOURNAL (YAKHTEH)[Internet]. 2008;10(SUPPLEMENT 1):48-48. Available from: https://sid.ir/paper/542430/en

IEEE: Copy

SH. EGHBAL SAEID, K. GHAEDI, MOHAMMAD HOSSEIN NASRESFAHANI, and N. AMIR MOZAFARI, “LIPOFECTAMINE DOES NOT ENHANCE BOVINE SPERM TRANSFECTION EFFICIENCY,” CELL JOURNAL (YAKHTEH), vol. 10, no. SUPPLEMENT 1, pp. 48–48, 2008, [Online]. Available: https://sid.ir/paper/542430/en

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