DNA and Lab Tools
What side is the 5 prime? The 3 prime?
5 prime is the phosphorus; 3' is the hydroxyll
how many ug are in 250 mg?
2.5*10^5 ug
What did we seperate out in the SEC lab?
Hemoglobin and B12
How many primers do you need? What ways do they run?
2; forward and reverse
5-->3'
3'-->5'
What does a project manager do?
Organize, create timelines, work with partners etc
What does the sugar backbone look like?
The oxygen ring
you are preparing 3g of agarose in 70 ml of buffer. What is the concentration of the gel?
4.3%
By size
How many loci did we use in our lab? How many are used in profiling?
1,13
What is the formula for error analysis?
Accepted-Found/Accepted *100
What is the difference between chromosomes, genes and DNA
Chromosomes are made of DNA and genes are segments of DNA
You have a 200x buffer and you want to make a 1x buffer that is 100 ml. How much of the 200x buffer will you need?
.5 ml
How do you know when a fraction will be included? excluded?
Explain why DNA fragments separate according to size in an electrophoresis gel?
Restriction enzymes cut into different sizes. DNA moves from negative to positive. Smaller fragments move farther than larger fragments which will move more down the gel
You grab a .2-200ul micropipette it reads
0
1
2
What is the reading?
12 ul
What molecules may interfere with DNA extraction?
DNAses
You have 2 1L serial dilutions with a factor of 70:30. How many ml of the original 100% stock solution is in the 2nd vial?
700 ml in the first
490 ml in the second :)
Samples travel faster or slower if they are bigger in SEC?
faster
What are the 3 steps of PCR with temperatures
Denaturing(94C), Anneal (55C) Extend (72 C)
Spell your teachers last name
How does the % agarose impact the bands of a gel?
As you increase % the bands will separate more. Need to ensure they aren't too far apart (that they fall off or that the gel is too solid) and also that the bands separate enough to be readable.
You have 3 serial dilutions with a factor of 90:10 at 70ml each. How much will you take from each solution and how much water will you need starting from a 95% solution?
1) 66.3 ml solution and 3.7 ml H20
2) 63 ml solution 7ml H20
3) 63 ml solution 7ml H20
Why do we drain the buffer?
so that the sample stays compact; purity; we don't want our sample to dilute
What determines where a restriction enzyme will cut?
a group of enzymes in bacteria, which when added to any DNA will result in the breakage [hydrolysis] of the sugar-phosphate bond between certain specific nucleotide bases [recognition sites].
This causes the double strand of DNA to break along the recognition site and the DNA molecule becomes fractured into two pieces. These molecular scissors or “cutting” enzymes are restriction endonucleases.
What are all of the parts of active listening? (5)
Open questions, summarizing, reflecting, clarifying, reacting