Biotechnology Lab I - Exam 2 4, (7 points) The next step is to analyze the genom
ID: 302566 • Letter: B
Question
Biotechnology Lab I - Exam 2 4, (7 points) The next step is to analyze the genomic DNA restriction digests on a 0.8% Summer 2018 agarose gel. When you checked the container with the IX TAE electrophoresis buffer, you found it was empty. So, you decided to make 10 liters of IX TAE Buffer using the lab stock of 50X TAE Buffer solution. But, you then found that the container with the 50X TAE Buffer was also empty. So, now you need to make 1,000ml of 50X TAE Electrophoresis Buffer. a. X TAE Buffer is 2mM EDTA in 40mM Tris with the pH adjusted to 8.0 with acetic acid. The formula weight for Tris is 121 g/mole. The formula weight for EDTA is 372 g/mole. To make 1,000ml of 50X TAE, you dissolved g Tris and g EDTA in water. After adjusting the pH to 8.0 with acetic acid, water was added to a final volume ml. You then made 20liters of IX TAR huffer by pouring ml 50X TAL nto an empty liters of water to bring the solution to a final volume ofjo liters. ml 1X container and then adding b. To make 400ml of 0.8% agarose, you combined TAE (ignore the volume occupied by the agarose) and t agarose. g agarose with heated the solution to boiling to melt the 2 mmExplanation / Answer
Answer A
242g of tris
18.61g of EDTA for
700ml
becasue add 242 g tris base and 18.61 g EDTA in ~700 ml Sterile ddH2O and stir it until completely dissolved, and then add 57.1 ml glacial acetic acid gradually, then adjust the volume up to 1L with ddH2O.
200 ml 50 x TAE
because for 1L : 20ml buffer + 980ml ddH2O for 50 ml of 1x solution
For 10L solution ; 200ml buffer + 9800ml dd H2O
Answer B
3.2g of agarose
because if you are making a 10mL gel that you want to be 0.8%, the amount of agarose to use is 0.08g, so..
for 400ml it would be 3.20g of agarose
400ml
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