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=== 1. QIAamp DNA Blood Mini Kit === : - Resuspend lyophilized QIAGEN Protease with 1.2 ml protease solvent. : - Use carrier DNA (e.g., poly dA, poly dT, poly dA:dT) when the sample is low-copy (i.e., when <10,000 copies are present) Starting material: 200ul for blood, plasma, serum or buffy coat ==== Procedures ==== : <u>Preparing lysate</u> :: - Pipet '''20 µl''' of '''QIAGEN Protease''' (or proteinase K) into the bottom of a 1.5 ml tube :: - Add '''200 µl sample''' to the tube. If the volume of sample less than 200ul, adjust volume with PBS :: - Add '''200 µl''' of '''Buffer AL''' to the sample. Mix by pulse-vortexing for 15 s :: - Incubate at 56°C for 10 min : <u>Binding DNA</u> :: - Briefly spin down the tube before adding 200ul '''100% EtOH'''. Mix again by pulse-vortexing for 15 s and briefly spin down :: - Transfer spnt to QIAamp Mini spin column & centrifuge at 6,000xg for 1 min : <u>Washing DNA</u> :: - Transfer QIAamp Mini spin column in a clean 2 ml collection tube :: - Add '''500 µl''' of '''Buffer AW1''' & centrifuge at 6,000xg for 1 min :: - Transfer QIAamp Mini spin column in a clean 2 ml collection tube :: - Add '''500 µl''' of '''Buffer AW2''' & centrifuge at 20,000xg for 3min : <u>Eluting DNA</u> :: - Transfer the QIAamp Mini spin column in a new 2 ml collection tube and centrifuge at 20,000xg for 1 min (full speed) :: - Place the QIAamp Mini spin column in a clean 1.5 ml tube & add appropriate volume of '''Buffer AE''' or distilled water (manual recommends 200ul). :: - Incubate at RT for 5 min & centrifuge at 6,000xg for 1 min <br>
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