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Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
Strand Cdna Synthesis Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
Gdna Eraser, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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TaKaRa prime scripttm rt pcr kit
Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
Prime Scripttm Rt Pcr Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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TaKaRa primescript rt master mix kit
Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
Primescript Rt Master Mix Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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TaKaRa cdna synthesis kit
Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
Cdna Synthesis Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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TaKaRa one step tb green primescript rt pcr kit ii
Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
One Step Tb Green Primescript Rt Pcr Kit Ii, supplied by TaKaRa, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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TaKaRa one step sybr prime script rt pcr kit
Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
One Step Sybr Prime Script Rt Pcr Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Yeasen Biotechnology two-step primescript mirna cdna synthesis kit
Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
Two Step Primescript Mirna Cdna Synthesis Kit, supplied by Yeasen Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Promega primescripttm reverse transcription (rt) reagent kit
Figure 6. Southern blot analysis of PCR products obtained from <t>total</t> <t>RNA</t> isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml <t>cDNA</t> mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.
Primescripttm Reverse Transcription (Rt) Reagent Kit, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Figure 6. Southern blot analysis of PCR products obtained from total RNA isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml cDNA mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.

Journal: Stem cells (Dayton, Ohio)

Article Title: Adenovirus mediated alpha interferon (IFN-alpha) gene transfer into CD34+ cells and CML mononuclear cells.

doi: 10.1002/stem.150386

Figure Lengend Snippet: Figure 6. Southern blot analysis of PCR products obtained from total RNA isolated from CD34+ cells transfected with AdCMV-IFN- α gene. Lane 1, PCR products amplified from nontransfected (control) CD34+ cell RNA; lane 2, RNA obtained from CD34+ cells transfected with AdCMV-IFN-α after 24 h; lanes 3 and 4, RNA from CFU-GM clones after 12 and 14 days of culture, respectively; lanes 5 and 6, RNA from BFU-E clones after 12 and 14 days of culture. Total RNA was extracted from CD34+ cells, BFU-E or CFU-GM clones. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml cDNA mixture of a total reaction volume of 25 ml. Aliquots of the reaction mixture (10 ml) were subjected to electrophoresis in a 0.8% agarose (Sigma; St. Louis, MO) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, (gift of Dr. Asano) using α-32d-CTP. Filters were exposed to x-ray for 48 h at –80°C.

Article Snippet: RNA is reverse-transcribed using the first strand cDNA synthesis kit from Clontech, Inc. (Palo Alto, CA).

Techniques: Southern Blot, Isolation, Transfection, Amplification, Control, Clone Assay, Electrophoresis

Figure 7. Southern blot analysis of PCR products obtained from total RNA isolated from CML hematopoietic stem cells (BMMNC) transfected with AdCMV-IFN-α gene. Lane 1, PCR products amplified from nontransfected (control) cell RNA; lane 2, RNA obtained from cells transfected with AdCMV-IFN-α after 24 h; lane 3, RNA from CFU-GM clones after 12 days; lane 4, CFU-GM clones after 14 days of culture; lane 5, no RNA was added; and lane 6, positive control. Total RNA was extracted from stem cells or CFU-GM clones by one single extraction. cDNA was synthesized from total RNA. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml cDNA mixture of a total reaction vol- ume of 25 ml. Aliquots of the reaction mixture (10 ml) were sub- jected to electrophoresis in a 0.8% agarose (Sigma) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, using α-32P-CTP. Filters were exposed to x-ray for 48 h at –80°C.

Journal: Stem cells (Dayton, Ohio)

Article Title: Adenovirus mediated alpha interferon (IFN-alpha) gene transfer into CD34+ cells and CML mononuclear cells.

doi: 10.1002/stem.150386

Figure Lengend Snippet: Figure 7. Southern blot analysis of PCR products obtained from total RNA isolated from CML hematopoietic stem cells (BMMNC) transfected with AdCMV-IFN-α gene. Lane 1, PCR products amplified from nontransfected (control) cell RNA; lane 2, RNA obtained from cells transfected with AdCMV-IFN-α after 24 h; lane 3, RNA from CFU-GM clones after 12 days; lane 4, CFU-GM clones after 14 days of culture; lane 5, no RNA was added; and lane 6, positive control. Total RNA was extracted from stem cells or CFU-GM clones by one single extraction. cDNA was synthesized from total RNA. IFN-α transcripts were amplified by PCR using 2 ml of 100 ml cDNA mixture of a total reaction vol- ume of 25 ml. Aliquots of the reaction mixture (10 ml) were sub- jected to electrophoresis in a 0.8% agarose (Sigma) gel. Nucleic acids were transferred to nitrocellulose by blotting and probed with IFN-α cDNA probes, using α-32P-CTP. Filters were exposed to x-ray for 48 h at –80°C.

Article Snippet: RNA is reverse-transcribed using the first strand cDNA synthesis kit from Clontech, Inc. (Palo Alto, CA).

Techniques: Southern Blot, Isolation, Transfection, Amplification, Control, Clone Assay, Positive Control, Extraction, Synthesized, Electrophoresis