May 05, 2022
Preparing Borax RapidBuffer for electrophoresis
- Jenny Molloy1,
- Nadine Mowoh2,
- Stephane Fadanka2,
- Cordellia Fulai2
- 1University of Cambridge, Beneficial Bio;
- 2Mboalab
- Beneficial BioTech. support email: [email protected]
Protocol Citation: Jenny Molloy, Nadine Mowoh, Stephane Fadanka, Cordellia Fulai 2022. Preparing Borax RapidBuffer for electrophoresis. protocols.io https://dx.doi.org/10.17504/protocols.io.ewov1nqopgr2/v1
License: This is an open access protocol 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
Protocol status: Working
We use this protocol and it’s working
Created: March 01, 2022
Last Modified: June 09, 2022
Protocol Integer ID: 58893
Keywords: borax rapidbuffer for electrophoresi, preparation of gel electrophoresis buffer, gel electrophoresis buffer, running dna gel, standard dna gel buffer, preparing borax rapidbuffer, dna gel, problems with gel integrity, separation of dna fragment, electrophoresi, 10mm sodium tetraborate, gel integrity, small gel, borax, cm gel, gel, conduction buffers like tbe, dna fragment, dna, high voltage
Abstract
Molecular biologists often use tris-based conduction buffers like TBE or TAE in running DNA gels. These buffers are known to overheat at high voltages, causing problems with gel integrity, sample denaturation and more. The voltage thus needs to be limited to a maximum of 5-10 volts/cm (e.g 100 volts for a 10 cm gel) leading to extended running time, sometimes to hours.
With 10mM Sodium Tetraborate (CAS#: 1330-43-4, Na2B4O7, Borax), the voltage can be increased to 35 volts/cm without any problem, reducing the time taken to run gels by up to 7 times for standard applications (e.g. separation of DNA fragments from 100 bp - 5 kbp).
Our lab’s standard DNA gel buffer has become 20 g of Borax into 1 l of water which gives a 20x stock solution. Small gels run in 10-15 minutes at 200V.
SCOPE:
Preparation of gel electrophoresis buffer for separation of DNA fragments from 100 bp - 5 kbp
Materials
Reagents
- Borax (Sodium Tetraborate, CAS#: 1330-43-4)
Materials and Equipment
- Beaker
- Measuring cylinder
- Distilled water
- Container with lid
- Weighing balance
- Weighing boat and spatula
- Magnetic stirrer
Troubleshooting
Preparing
Weigh out a certain amount of borate and place into a vacuum Tupperware half filled with silica beads.
place the lid on top and cover tightly, then use the vacuum pump to extract air inside and allow to dry for12:00:00 at 37 °C ).
1.2 Carefully pour it out into zip lock bags and seal to avoid it from absorbing moisture and store in cool dry place.
12h
Preparation of 10x Borax RapidBuffer Solution
10m
Use a spatula to measure out and weigh 10 g of Borax (Sodium Tetraborate, CAS#: 1330-43-4)
Pour powder into a graduated 1000 mL glass beaker or flask
Using a 1000 mL measuring cylinder, measure800 mL of distilled or deionised water and add to the 1000 mL beaker.
Add a magnetic flea and use a magnetic stirrer at Room temperature to fully dissolve the borax powder.
2m
Add distilled or de-ionised water to a total of 1000 mL and stir again for 00:03:00
3m
Pour the prepared solution into a clean, labelled screw-top 1000 mL glass or plastic storage bottle and store at Room temperature until use.
Preparation of 1x Borax RapidBuffer Solution (500ml)
10m
Use a 100 mL measuring cylinder to measure out 50 mL of 10X RapidBuffer into a 500 mL graduated beaker or flask.
Add approx 450 mL distilled or deionised water, sufficient to make up the volume to 500 mL
1X RapidBuffer solution is now ready for use and can be stored at Room temperature for use within one month.