Mouse tissue lysis for genotyping protocol

Solutions/reagents:  Taq DNA polymerase 10X buffer with MgCl2  (diluted 1:10 in ddH2)  proteinase K  (20 mg/ml) tail or tissue chunk Equipment: IncubatorCentrifuge Steps: <ol>  Tissue lysis to release DNA  Measure out <font color=#357EC7>100 µl  of <a href="#Taq DNA polymerase 10X buffer with MgCl2" ><font color=#357EC7>Taq DNA polymerase 10X buffer with MgCl2 </a> into tail or tissue chunk. Add <font color=#357EC7>2 µl  of <a href="#proteinase K" ><font color=#357EC7>proteinase K </a>. </li> Incubate  at <font color=#357EC7>50°C  for <font color=#357EC7>3 - 12 hrs(overnight)  with shaking at 300 rpm. <font color = "#800517">proteinase K is stable over a broad pH range (4.0 to 12.5, optimum pH 8.0) and is also stable over the temperature range of 25 to 65°C. </li>  Proteinase K inactivation  Incubate  at <font color=#357EC7>95°C  for <font color=#357EC7>10 - 20 mins  with shaking at 300 rpm. Centrifuge at a speed of <font color=#357EC7>13000 rpm for <font color=#357EC7>3 mins  at <font color=#357EC7><b><font color=#357EC7>room temperature  </b> and aspirate out <font color=#357EC7>80 µl  of top layer into sterile 1.5-ml microcentrifuge tube (2). Discard bottom layer. </li>  Storage before PCR  NOTE: If sample is/are used within <font color=#357EC7>1 hr , set temperature to <font color=#357EC7>4°C  and use, else store it at <font color=#357EC7>-20°C .</li> </ol>