Difference between revisions of "PCR Analysis of Tail DNA"

Latest revision as of 17:00, 8 June 2020

see Genotyping Program for strain specific details

First, make 250ul of 0.4um working stock primers from 100um Primary Stocks.

Use the following Volumes per 25ul Reaction:

Per sample (1X)

Run "specfic" PCR Program for gene of interest (approx 2 hours).

see Preparing an Agarose Gel for details on preparing a DNA gel

@@ Line 1: / Line 1: @@
-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==
-Use the following Volumes per Reaction:
+First, make 250ul of 0.4um working stock primers from 100um Primary Stocks.
+#248 ul ddH20
+#1ul forward primer (100um)
+#1ul reverse primer (100um)
-#Buffer: 4 uL of 5X Go-Taq buffer ("Molecular Biology Stuff" box in freezer)
-#Forward Primer: 0.4ul
-#Reverse Primer: 0.4ul
-#dNTPs: 0.4uL 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)
-#Template: 1 uL
-Run PCR Program (approx 2 hours).
+Use the following Volumes per 25ul Reaction:
-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
+Per sample (1X)
+#Dream Tag Master mix: 12.5uL ("Molecular Biology Stuff" box in freezer)
+#0.4um Primer Mix: 5ul
+#Sterile ddH2O: 7.5ul
+*Template: 1 uL
+Run "specfic" PCR Program for gene of interest (approx 2 hours).
+*[[Genotyping Program]]
+*[[PCR Amplification of DNA]]
 see [[Preparing an Agarose Gel]] for details on preparing a DNA gel