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=Urea Gel Quantification= [[Daniel Jacobsen|Back to Main]] ==Protocol 1== 1. Prerun gel for at least 20 minutes at 250 Volts 2. Prepare samples, typical setup: {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> |- style="font-size:12pt" align="center" | align="center" width="65" height="35" | | width="65" | uL sample | width="65" | uL ladder | width="65" | uL TBE | width="65" | uL 2X urea buffer | width="65" | uL loaded |- style="font-size:12pt" align="center" | height="15" | samples | align="center" | 0.5 | align="center" | 0 | align="center" | 5.5 | align="center" | 6 | align="center" | 12 |- style="font-size:12pt" align="center" | height="15" | ladder 1 | align="center" | 0 | align="center" | 0.5 | align="center" | 5.5 | align="center" | 6 | align="center" | 12 |- style="font-size:12pt" align="center" | height="15" | ladder 2 | align="center" | 0 | align="center" | 1 | align="center" | 5 | align="center" | 6 | align="center" | 12 |} 3. Denature samples for 8 minutes at 65C 4. Snap cool using freezer box 5. Load samples onto gel and run for 20-25 minutes at 250V 6. Stain with 3 uL SYBR gold for at least 15 minutes ===Example Gel Image Using Protocol 1=== [[Image:LambdaExo-030613.jpgβ|600px]] ==Protocol 2== 1. Preheat TBE buffer by incubating it in 60C incubator Microwave to do it quickly, but make sure not to get the buffer too hot (ideal: 45-60C) 2. Prerun gel in heated buffer at 250V for ~15-20 minutes 3. Prepare samples according to following table: {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> |- style="font-size:12pt" align="center" | align="center" width="65" height="35" | | width="65" | uL sample | width="65" | uL ladder | width="65" | uL TBE | width="65" | uL 2X urea buffer | width="65" | uL loaded |- style="font-size:12pt" align="center" | height="15" | samples | align="center" | 0.5 | align="center" | 0 | align="center" | 5.5 | align="center" | 6 | align="center" | 12 |- style="font-size:12pt" align="center" | height="15" | ladder 1 | align="center" | 0 | align="center" | 0.5 | align="center" | 5.5 | align="center" | 6 | align="center" | 12 |- style="font-size:12pt" align="center" | height="15" | ladder 2 | align="center" | 0 | align="center" | 1 | align="center" | 5 | align="center" | 6 | align="center" | 12 |} 4. Denature samples for 8 minutes at 65C 5. Load samples and run gel for 20-25 minutes at 65C 6. Stain with 3 uL SYBR gold for at least 15 minutes (rec: 20-25) ===Example Gel Image using Protocol 2=== [[Image:PrimerQuant_050813.jpg|600px]] ==Invitrogen Low Mass Ladder== Invitrogen, Catalog #: 10068-013, $113.00 Infosheet: [[File:InvitrogenLowMassLadder.pdfβ]] [[Image:LowMassLadder_Invitrogen.png|600px]]
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