Gateway Cloning: Simple and Multisite Gateway Cloning - LR and BP Cloning

[@albasiquier516]

Thank you so much for this video, it was very helpful!! Your explanations are clear and very well structured, what makes them very easy to follow. I also love your handwriting and drawings, they look very good! Thanks for helping me prepare for my Genetics Engeneering midterm!! :)

[@roomiemcgee8899]

This was so helpful. Thank you! Preparing to transfect drosophila for the first time.

[@meadowbeeflying]

Better guide to cloning is not exist :)

[@Rajat_1010]

Thank you for uploading this!!

[@김대빈-w5m]

10:05 the attL site and attR site... the word L and R do not come from Left and Right, right? If they are, how does it suddenly come with two attL sites in one Vector and two attR sites in another? at 05:38 they were mixed as L R and R L after B B' , P P' recombination.. So I think R-DNAi-L and L-ccdB-R recombination will result back in P-ccdB-P and B-DNAi-B right? And also after LR recombination 12:26 the names attB1, attB3, attB2 are just randomly given? So it is also fine to name them attP1, attP3 and attP2?

[@theCrux]

At 3:54 the L/R do come from Left and Right. At 5:38 the custom vectors contain B B and P P. At 9:57 the custom vectors contain L L and R R. At 12:26 the names are not truly random. There are attP and attB pairs called attP1/attB1, attP2/attB2 etc. Theirs products are likewise called attL1/attR1, attL2/attR2, etc.

[@김대빈-w5m]

@@theCrux Thank you for you kind explanation! Sorry I think I confused you... What I meant was 3:11 you marked a half block as B and B' and also P and P'. At 4:52 you wrote B B and P P but just as hole block each. So my questions is the name B B and P P as block just mad up by yourself? Another question is, at 5:38 you wrote L and R in the donor vector (L-ccdB-R) just as you described at 4:01 (attL-virus DNA-attR). They have some combination (B+P'=L and P+B'=R) given that the whole block of B and P are divided into B and B' / P and P'. So at the end there are L-ccd-R part in the donor vector and R-DNAi-L part in the final vector at 5:38. But how come you write two vectors with (L L and R R) at 10:09? Becuase after B P reaction the Donors include either R+L or L+R (05:40). OR it is just made up by yourself to exlain LR reaction so it results in the final product as B B and P P? So it has nothing to do with previous stories of lytic phase or the initial BP reaction because after BP reaction the vectors have either L+R or R+L... I hope you understand my point. Thank you for your answer in advance!