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Second strands of driver and tester DNA were synthesized using T4 DNA polymerase and restriction digested by RsaI to create blunt-ended double stranded cDNA for subtraction.
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Tester DNA was then melted and reannealed with an excess of driver DNA and the remaining single strand driver DNA (normalized or subtracted library) was then purified by hydroxyapatite chromatography.
Hybridization of the anti-sense strands from tester cDNA and sense strands from driver cDNA was carried out in 1× hybridization buffer (50 mM Tris-HCl pH 8.0, 0.5 M NaCl, 0.2 mM EDTA) at 68°C for 6 h.
Dyfed-Powys Police said it had received dozens of calls, including of roads becoming impassable due to flash floods and of a stranded driver.
Volunteers were praised for rallying to the aid of stranded drivers in Kent and Sussex, with farmers and builders mobilising tractors and diggers to clear minor roads and 30 members of a Plymouth Brethren church working shifts with seven 4x4s to bring medical supplies, hot drinks and reassurance to travellers stuck on the A23.
Dyfed-Powys Police received dozens of calls, including reports of roads becoming impassable due to flash floods and one of a stranded driver.
Those tow drivers do have to download and install a native mobile app, but once they're within range of a stranded driver they'll respond to that and provide two things — their ETA to the car's location and a quick price quote.
Hundreds of drivers were stranded in their cars overnight and some schools are closed.
Hundreds of schools were still shut in Wales and dozens of drivers were stranded in their vehicles.
Heavy snow has been causing travel problems after hundreds of drivers were stranded in their cars overnight in southern England.
Heavy snow is causing travel problems for a second day after hundreds of drivers were stranded in their cars overnight in southern England.
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