PCR Analysis of Tail DNA: Difference between revisions

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-see [[Genotyping Details]] for strain specific details
+see [[Genotyping Program]] for strain specific details
 ==Materials==
+# Dream Taq Green master mix
+# Specific gene Primers (0.4um Working stock)
+# Tail digest DNA
+# ddH2O
 ==Protocol==
+First, make 250ul of 0.4um working stock primers from 100um Primary Stocks.
+#248 ul ddH20
+#1ul forward primer (100um)
+#1ul reverse primer (100um)
-Use the following Volumes per Reaction:
-Buffer: 4 uL of 5X Go-Taq buffer ("Molecular Biology Stuff" box in freezer)
+Use the following Volumes per 25ul Reaction:
-Forward Primer: .4ul
+Per sample (1X)
+#Dream Tag Master mix: 12.5uL ("Molecular Biology Stuff" box in freezer)
+#0.4um Primer Mix: 5ul
+#Sterile ddH2O: 7.5ul
-Reverse Primer: .4ul
+*Template: 1 uL
-dNTPs: .4uL of 2 mM ("Molecular Biology Stuff" box in freezer)
-Sterile water: 13.6 uL
-Polymerase Go-Taq: 1 uL (6th floor in Genotype Yellow Box in freezer)
+Run "specfic" PCR Program for gene of interest (approx 2 hours).
-Template: 1 uL
+*[[Genotyping Program]]
+*[[PCR Amplification of DNA]]
+see [[Preparing an Agarose Gel]] for details on preparing a DNA gel
+[[Category: Genotyping]]
-Run PCR Program (approx 2 hours).
+[[Category: Mouse Work]]
-Use Cycler 1 on 6th Floor
-Login: Sergey, Just press enter to Login
-Under Genotype folder, pick Ingles program for Ingles genotyping
-Under Genotype folder, pick regpcr program for PLT genotyping
-Make sure to press enter 2x once to confirm Tubes and second time to start PCR
-see [[Preparing an Agarose Gel]] for details on preparing a DNA gel