Week 3: How to make 1% Gel for PCR

 Week 3: Making 1% Gel for PCR


    In lab this week, I learned and experimented with 1% gel and how to make it. It requires some memorization on measuring and materials. It is not difficult but it just has a lot of steps.

Materials Needed:
- TAE Buffer (Key Ingredient)

- 30 ml Electrophoresis Machine

- 300 ml OR 0.3 grams of Agarose

Process:

Measuring Agarose- 30 mL x 1% (0.01)= 300 ml OR 0.3 grams

1. Put 300 ml/30g of agarose into a microwavable beaker of 30ml TAE buffer and swirl to mix. 

2. Make a stopper with rolls of paper towels and inserting in the beaker's mouth (making sure it is all SOAKED and it fits TIGHTLY in the beaker's mouth)

3. Put the beaker in a microwave for 30 seconds. If liquid is not clear, add 30 seconds more) 

4. Wait for mixture to cool since it is extremely hot and almost at boiling temperature. 
       ---> This prevents the Electrophoresis Machine's plastic to be warped when pouring the gel substance when it is still extremely hot.

5. Prepare the Electrophoresis Machine. 
        ---> Negative charge going towards the positive charge so the ladder must go near the negative charge. Insert the black stoppers tightly between the center's left and right to avoid gel substance to overflow onto the side. 

6. After beaker has cooled down (Is able to hold the beaker for more than 10 seconds with bare hands), gently and slowly pour in the gel substance from the CORNER. 

7. Check if the gel substance is overflowing. If not, wait about 10-20 minutes for gel to harden. 

8. Once gel is harden, slowly remove stoppers and ladder. Fill up to the MAX with TAE buffer starting in the ladders to prevent air-bubbles. 

9. Gel is now ready for the PCR and UV Dye to be inserted.

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