Lecture 7

Question 1

Why do we need to clean the DNA?

Answer 1

To keep the desired DNA size (300 bp)
-Larger and smaller DNA sizes will be washed off

Question 2

How are beads able to bind to different sizes of DNA

Answer 2

The buffer volume dictates the size
-the pieces of DNA w/ the correct size stays bound to the beads everything else gets washed off/ is in the supernatant

Question 3

After tagmentation, can Qubit measure a higher DNA concentration than the one you measured before tagmentation?

Answer 3

Yes
-We expect it to be higher after tagmentation b/c we amplified the DNA
-if it is lower–>we lost DNA during the process

Question 4

What is Tape Station Analysis (QC)?

Answer 4

DNA samples are checked via high quality electrophoresis for quality
-Tells average size of DNA fragments in sample

Question 5

If you measure your sample’s DNA concentration by Qubit and Nanodrop, can Qubit measure a higher DNA concentration than Nanodrop?

Answer 5

No
-Qubit ONLY measures dsDNA (intact DNA)
-Nanodrop measures everything (degraded DNA, RNA, etc)
-Therefore, Qubit conc is lower than Nanodrop

Question 6

True or false: Clusters of similar fragments allows for fluorescent signal

Answer 6

True
-clusters allow us to see fluorescence b/c they produce enough of the signal