Daniel:Protocols/TBUgel

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Urea Gel Quantification[edit]

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Protocol 1[edit]

1. Prerun gel for at least 20 minutes at 250 Volts
2. Prepare samples, typical setup:
  uL sample uL ladder uL TBE uL 2X urea buffer uL loaded
samples 0.5 0 5.5 6 12
ladder 1 0 0.5 5.5 6 12
ladder 2 0 1 5 6 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[edit]

File:LambdaExo-030613.jpg

Protocol 2[edit]

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:
  uL sample uL ladder uL TBE uL 2X urea buffer uL loaded
samples 0.5 0 5.5 6 12
ladder 1 0 0.5 5.5 6 12
ladder 2 0 1 5 6 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[edit]

File:PrimerQuant 050813.jpg

Invitrogen Low Mass Ladder[edit]

Invitrogen, Catalog #: 10068-013, $113.00

Infosheet: File:InvitrogenLowMassLadder.pdf

File:LowMassLadder Invitrogen.png