Simply Cloning - Chapter 4 - Gel Purification

[@CloningStrategies]

It has to do with DNA supercoiling. When plasmid is circular (uncut vector lane) the supercoils make it more compact and it moves through the gel more easily. Also the fragment that is removed is very small, about 40 base pairs.

[@SharathChandraRaoKV]

Thanks so much....i found them very useful and handy..

[@nemoco99]

Hi Andriy, great video! Have you tried purification with chemical or classic methods instead of purification kits, I need to do a blunt end ligation but I need to purificate without kit. Thank you.

[@yaskoo]

Great videos ...thanks ..

[@SharathChandraRaoKV]

Hi Andriy, Could you please send the protocols or the link to get the cloning experiments protocols. Your tutorial videos are awesome....Thanks for the videos...

[@CloningStrategies]

Hi Sharath, the prorocols are here: www.cloningstrategies.com/protocols.pdf

[@MrSpeedyAce]

Why does the cut vector show up as bigger DNA than the uncut? The cut has a chunk that is missing from it so shouldn't it come up as a lower band than the uncut?

[@MrCoolio1985]

Great thank you

[@machshung2615]

Tks ad

[@MrSpeedyAce]

@CloningStrategies Thanks for the reply :-)

[@mrphysh]

The digest of the PSAT6mcs cuts out a little piece of the plasmid and makes it linear. This little piece might be seen in the gel electrophoresis, but it is not. Am I understanding this correctly?

[@mrphysh]

Now I see it, The 40 base piece just disappears into the solvent front ....(If I may use chromatography language)