Oct 23, 2020
PREPARING 6X NEB LOADING BUFFER CONTAINING GELGREEN (GG) labeled ‘6XNEB+GG’
This document is a draft, published without a DOI.
- 1UCSC
- UCSC BME 22L
Document Citation: Ikenna Anigbogu 2020. PREPARING 6X NEB LOADING BUFFER CONTAINING GELGREEN (GG) labeled ‘6XNEB+GG’. protocols.io https://protocols.io/view/preparing-6x-neb-loading-buffer-containing-gelgree-bnv5me86
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 23, 2020
Last Modified: October 23, 2020
Document Integer ID: 43677
Disclaimer
DISCLAIMER – FOR INFORMATIONAL PURPOSES ONLY; USE AT YOUR OWN RISK
The protocol content here is for informational purposes only and does not constitute legal, medical, clinical, or safety advice, or otherwise; content added to protocols.io is not peer reviewed and may not have undergone a formal approval of any kind. Information presented in this protocol should not substitute for independent professional judgment, advice, diagnosis, or treatment. Any action you take or refrain from taking using or relying upon the information presented here is strictly at your own risk. You agree that neither the Company nor any of the authors, contributors, administrators, or anyone else associated with protocols.io, can be held responsible for your use of the information contained in or linked to this protocol or any of our Sites/Apps and Services.
PREPARING 6X NEB LOADING BUFFER CONTAINING GELGREEN (GG) labeled ‘6XNEB+GG’
23 OCTOBER 2020
DRAFT
OBJECTIVE
Prepare 6X NEB Buffer + Gelgreen that will be used in your DNA gel electrophoresis experiments. In our research lab, we add 6 uL of Gelgreen stock solution to 1 mL of 6X NEB Buffer to get the working mixture. You will be preparing a smaller solution so you will need to undertake an extra dilution step.
METHOD
1) Using your 20 uL pipette, add 2 uL of Gelgreen (small tube of red solution in your recent UPS packet labeled ‘GG’) to a high quality 1.5 mL Eppendorf tube. Label the tube ‘GG 1:10’. (Note: Be careful when pipetting these small volumes. Practice by pipetting 2 uL of water before pipetting the Gelgreen.)
2) Also using your 20 uL pipette, add 18 uL of 6X NEB buffer (also in your recent UPS packet) to the GG 1:10 tube. (Note: 6X NEB buffer contains glycerol, so it can stick to the outside of the pipet tip). Store this tube at room temperature in the dark.
3) Using the 20 uL pipet, add 2.5 uL of the 1:10 GG working stock to a new high quality 1.5 mL Eppendorf tube. Label this tube 6XNEB+GG.
5) Add 39 uL of NEB 6X Loading buffer to the 6XNEB+GG tube. This is the solution you will use for your gels. Store the tube at room temperature in the dark.