Oct 20, 2020
Lab Notebook Template
This document is a draft, published without a DOI.
- 1UCSC
- UCSC BME 22L
Document Citation: Ikenna Anigbogu 2020. Lab Notebook Template. protocols.io https://protocols.io/view/lab-notebook-template-bnnpmddn
License: This is an open access document distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Created: October 20, 2020
Last Modified: October 20, 2020
Document Integer ID: 43439
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GEL ELECTROPHORESIS 2 : 1 KB GEL-BASED ASSAY
OBJECTIVES
- To visualize a 1 kilobase DNA ladder using a 1% agarose/0.5X TBE gel using Bento Lab.
- Test DNA staining by Gelgreen when caste in the gel using different amounts of DNA
- Practice gel electrophoresis skills.
- Practice laboratory notebook entries.
METHODS
- Casting a 1% agarose gel.
- Follow protocol "Agarose Gel Protocol " written by Dr. Akeson, with the following exceptions if any
2. Preparing the 1 kb DNA ladder samples. Samples are 0.5, 1.0, 2.0, 5.0 ug of NEB ladder. Combine as follows in cheap Eppendorf tubes.
| ug DNA | ul NEB ladder | ul 6X loading buffer | water ul | total volume (uL) | |
| 0.5 | 1 | 2 | 9 | 12 | |
| 1.0 | 2 | 2 | 8 | 12 | |
| 2.0 | 4 | 2 | 6 | 12 | |
| 5.0 | 10 | 2 | 0 | 12 |
3. Preparing the gel (See Dr. Akeson's gel protocol noted in 1 above)
4. Loading the sample
- Gel was placed on the Bento Box transilluminator taking care not to spill running buffer. Sample was loaded into lanes 3, 4, 5, and 6.
5. Running the gel.
- Standard Bento Lab protocol, i.e. 50V for one hour
RESULTS
1. Gel was visualized at 20 minutes. All lanes visible. 1.0 to 5.0 ug clearly overloaded.
CONCLUSION
- 0.5 ug lane was fine but not superb. Larger bands diffuse.
- Loading 1 ug or more DNA caused severe streaking due to lane overload