B) Restriction Digest Reactions-the buffer used a restriction digest is called C
ID: 185966 • Letter: B
Question
B) Restriction Digest Reactions-the buffer used a restriction digest is called Cutsmart Buffer. Cutsmart Buffer is provided by digest reaction contains 1xCutSmart Buffer, 2L phage (this volume is not included in your calculations), and sterile water. How much 10X CutSmart Buffer an sterile water should you add to the reaction? the company the makes restriction enzymes at a 10X concentration. A restriction DNA, a negligible amount of restriction enzyme 2HL DNA HL 10X CutSmart Buffer L sterile water 40L total volume How would your answer change if your restriction digest had a total volume of 25L rather than 40L? 2uL DNA HL 10X CutSmart Buffer L sterile water 25L total volume
Explanation / Answer
concentration of restriction enzyme is 10X
working concentration is 1X
therefore we require to dilute the cutsmart buffer 10/1= 10 times
for 40 l. reaction volume, cutsmart buffer required will be 40/ 10= 4l.
Total reaction for 40l is
2l DNA
4l cutsmart buffer
34l of sterile water
If the reaction volume is 25l, then the reaction volume will be
2l DNA
25/ 10= 2.5 l of cutsmart buffer and
20.5l of sterile water.
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