Sunday, May 2, 2021

ProteoCool Pills#4: Replacement of Ethidium bromide with a safe stain

 

Ethidium bromide thanks to its ability to show an intense fluorescence (ex 300nm/em 620nm) after binding with DNA.

 Ethidium bromide (EB) is inexpensive, sufficiently sensitive and very stable and thanks to this property has been the stain of choice for nucleic acid gel staining for decades.

 However on the other hands, EB is also a known powerful mutagen.

Recent years alternative reagents able to show an intense fluorescence after binding with DNA but not able to permeate cell membrane were developed:

 

My preferred one:

 

Gel REd - Biotium  (VWR cod. 41003) 

   https://biotium.com/product/gelred-nucleic-acid-gel-stain/

 

Safety assessment available at:

 

https://biotium.com/wp-content/uploads/2013/07/GelRed-and-GelGreen-Safety-Report.pdf

 

Cost: About 90€ for 500ul (stock 10000X) à  90€ for 5liter of agarose à 0,9€ for a mini gel (15x2well)

                                                                                                                                  2,5€ for a midi gel (up to 30x2 well)

                                                                                                                                     

Another alternative that I used in the past is the Sybr Safe (thermo cod. S33102) 

https://www.thermofisher.com/order/catalog/product/S33102?SID=srch-srp-S33102#/S33102?SID=srch-srp-S33102 

is another possible alternative. but in my experience, it shows limited stable under UV excitation.

 

More details are available in the following link

 

https://www.blogger.com/video.g?token=AD6v5dxB5wweSgHEks3TU3bm1QoGEYZU6GT8788soF0i7SEVvaQwM2AgHXmDY62fIzukFrMMDbFQEFOPMfSyV4GwvJHPY9U__KiUhmaCsx6NAwneL2yJYbaSu_DV-H4_to6BznMtlE59

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