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  3. 3 set up for the pcr tubes for a obtain 1 pcr tube

3 Set up for the PCR tubes for a Obtain 1 PCR tube containing Ready...

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3 Set up for the PCR tubes for a Obtain 1 PCR tube containing Ready To Go PCR beads for each sample three in total and label with the sample type group color and tubulin b Add 22 5 uL of the tubulin primer loading dye mix to each tube Allow several min for bead to dissolve c Add 2 5 L of each DNA sample to its respective tube and label 4 Close each tube and mix by tapping the bottom of the tube Keep tubes on ice until all groups are ready for the next step 5 Give your tubes to your TA to place into the Thermocycler The amplification run setup is as follows PCR Cycle Program 1 cycle 95 C 5 min 36 cycles 94 C 10 sec 55 C 5 sec 72 C 5 sec 1 cycle 72 C 1 min The PCR cycle includes a denaturation step 94 C a reannealing step 55 C and an elongation step 72 C PCR Beads contain PuReTaq DNA Polymerase 10 mM Tris HCl 50 mM KCl 1 5 mM MgCl2 200 M of each dNTP deoxyribunucleated triphosphorylated DNA bases Preparing for Gel Electrophoresis Make the following solution 75 ml 10x TBE Tris base Boric acid EDTA 0 9 M Tris 0 889 M boric acid 20 mM EDTA This solution will be used next week for the gel electrophoresis in Part C of the protocol

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