“Having had the green light to run all protocols and using
all four bacteria cultures I did exactly that and was left drained at the end
of that day” – Thursday, March 23rd
This week was dedicated to analyzing the products of the DNA
extraction methods using microvolume
UV-Vis spectrophotometer technology. The results are on the tables below and
can be overwhelming to look at.
There
were four culture broths used, each with a different color cap:
-Green
(g)
-White
(w)
-Red
(r)
-Blue
(b).
There
was a total of six protocols:
-Boiling
(BP)
-TE
Boiling (tBP)
-TE
Freeze and Thaw (tFT)
-Lysis
(L)
-Meat
Tenderizer (MT)
-Commercial
Kit (C)
Each
culture was used to conduct all of the protocols to determine which protocol was
the best. Given that there were four cultures, the experiment was conducted four
different times technically. Here are the results:
|
Sample ID
|
Nucleic Acid
|
Unit
|
260/280
|
Sample Type
|
Factor
|
|
BPg(Blank)
|
|||||
|
BPg
|
33.7
|
ng/µl
|
1.48
|
DNA
|
50
|
|
BPg
|
33.3
|
ng/µl
|
1.48
|
DNA
|
50
|
|
BPg
|
34
|
ng/µl
|
1.49
|
DNA
|
50
|
|
BPg
|
34
|
ng/µl
|
1.49
|
DNA
|
50
|
|
BPg
|
33.7
|
ng/µl
|
1.49
|
DNA
|
50
|
|
BPw(Reblank)
|
|||||
|
BPw
|
31.2
|
ng/µl
|
1.52
|
DNA
|
50
|
|
BPw
|
30.5
|
ng/µl
|
1.49
|
DNA
|
50
|
|
BPw
|
35.5
|
ng/µl
|
1.4
|
DNA
|
50
|
|
BPw
|
30.8
|
ng/µl
|
1.49
|
DNA
|
50
|
|
BPw
|
31.9
|
ng/µl
|
1.52
|
DNA
|
50
|
|
BPr(Reblank)
|
|||||
|
BPr
|
77.2
|
ng/µl
|
1.57
|
DNA
|
50
|
|
BPr
|
76.7
|
ng/µl
|
1.56
|
DNA
|
50
|
|
BPr
|
77.2
|
ng/µl
|
1.57
|
DNA
|
50
|
|
BPr
|
76.7
|
ng/µl
|
1.57
|
DNA
|
50
|
|
BPr
|
80.2
|
ng/µl
|
1.57
|
DNA
|
50
|
|
PBb(Reblank)
|
|||||
|
BPb
|
56.6
|
ng/µl
|
1.6
|
DNA
|
50
|
|
BPb
|
57.7
|
ng/µl
|
1.59
|
DNA
|
50
|
|
BPb
|
59
|
ng/µl
|
1.61
|
DNA
|
50
|
|
BPb
|
57.8
|
ng/µl
|
1.58
|
DNA
|
50
|
|
BPb
|
57.4
|
ng/µl
|
1.61
|
DNA
|
50
|
|
Sample ID
|
Nucleic Acid
|
Unit
|
260/280
|
Sample Type
|
Factor
|
|
tBPg(Blank)
|
|||||
|
tBPg
|
885.5
|
ng/µl
|
1.43
|
DNA
|
50
|
|
tBPg
|
883.3
|
ng/µl
|
1.44
|
DNA
|
50
|
|
tBPg
|
968
|
ng/µl
|
1.43
|
DNA
|
50
|
|
tBPg
|
885
|
ng/µl
|
1.44
|
DNA
|
50
|
|
tBPg
|
873.8
|
ng/µl
|
1.44
|
DNA
|
50
|
|
tBPw(Reblank)
|
|||||
|
tBPw
|
933.8
|
ng/µl
|
1.5
|
DNA
|
50
|
|
tBPw
|
956.7
|
ng/µl
|
1.5
|
DNA
|
50
|
|
tBPw
|
949.1
|
ng/µl
|
1.5
|
DNA
|
50
|
|
tBPw
|
954.4
|
ng/µl
|
1.5
|
DNA
|
50
|
|
tBPw
|
948.2
|
ng/µl
|
1.49
|
DNA
|
50
|
|
tBPr(Reblank)
|
|||||
|
tBPr
|
963
|
ng/µl
|
1.56
|
DNA
|
50
|
|
tBPr
|
963.8
|
ng/µl
|
1.56
|
DNA
|
50
|
|
tBPr
|
967.9
|
ng/µl
|
1.56
|
DNA
|
50
|
|
tBPr
|
968.6
|
ng/µl
|
1.56
|
DNA
|
50
|
|
tBPr
|
971.5
|
ng/µl
|
1.55
|
DNA
|
50
|
|
tBPb(Reblank)
|
|||||
|
tBPb
|
977.2
|
ng/µl
|
1.58
|
DNA
|
50
|
|
tBPb
|
997.7
|
ng/µl
|
1.57
|
DNA
|
50
|
|
tBPb
|
987.7
|
ng/µl
|
1.57
|
DNA
|
50
|
|
tBPb
|
966.8
|
ng/µl
|
1.57
|
DNA
|
50
|
|
tBPb
|
993.3
|
ng/µl
|
1.57
|
DNA
|
50
|
|
Sample ID
|
Nucleic Acid
|
Unit
|
260/280
|
Sample Type
|
Factor
|
|
tFTg(Blank)
|
|||||
|
tFTg
|
212.4
|
ng/µl
|
1.62
|
DNA
|
50
|
|
tFTg
|
212.1
|
ng/µl
|
1.63
|
DNA
|
50
|
|
tFTg
|
206.4
|
ng/µl
|
1.58
|
DNA
|
50
|
|
tFTg
|
209.1
|
ng/µl
|
1.64
|
DNA
|
50
|
|
tFTg
|
217.5
|
ng/µl
|
1.64
|
DNA
|
50
|
|
tFTw(Reblank)
|
|||||
|
tFTw
|
253.2
|
ng/µl
|
1.54
|
DNA
|
50
|
|
tFTw
|
249.3
|
ng/µl
|
1.54
|
DNA
|
50
|
|
tFTw
|
253.2
|
ng/µl
|
1.54
|
DNA
|
50
|
|
tFTw
|
256.1
|
ng/µl
|
1.54
|
DNA
|
50
|
|
tFTw
|
254.5
|
ng/µl
|
1.54
|
DNA
|
50
|
|
tFTr(Reblank)
|
|||||
|
tFTr
|
170.8
|
ng/µl
|
1.53
|
DNA
|
50
|
|
tFTr
|
171.8
|
ng/µl
|
1.53
|
DNA
|
50
|
|
tFTr
|
174.2
|
ng/µl
|
1.54
|
DNA
|
50
|
|
tFTr
|
171.7
|
ng/µl
|
1.53
|
DNA
|
50
|
|
tFTr
|
167.6
|
ng/µl
|
1.49
|
DNA
|
50
|
|
tFTb(Reblank)
|
|||||
|
tFTb
|
172.6
|
ng/µl
|
1.58
|
DNA
|
50
|
|
tFTb
|
176.9
|
ng/µl
|
1.58
|
DNA
|
50
|
|
tFTb
|
175.2
|
ng/µl
|
1.57
|
DNA
|
50
|
|
tFTb
|
173.8
|
ng/µl
|
1.56
|
DNA
|
50
|
|
tFTb
|
175.2
|
ng/µl
|
1.57
|
DNA
|
50
|
|
Sample ID
|
Nucleic Acid
|
Unit
|
260/280
|
Sample Type
|
Factor
|
|
Lg(Blank)
|
|||||
|
Lg
|
258.6
|
ng/µl
|
1.68
|
DNA
|
50
|
|
Lg
|
356.1
|
ng/µl
|
1.71
|
DNA
|
50
|
|
Lg
|
186.4
|
ng/µl
|
1.58
|
DNA
|
50
|
|
Lg
|
301.7
|
ng/µl
|
1.63
|
DNA
|
50
|
|
Lg
|
112.8
|
ng/µl
|
1.69
|
DNA
|
50
|
|
Lw(Reblank)
|
|||||
|
Lw
|
261.9
|
ng/µl
|
1.57
|
DNA
|
50
|
|
Lw
|
263.2
|
ng/µl
|
1.57
|
DNA
|
50
|
|
Lw
|
317.3
|
ng/µl
|
1.57
|
DNA
|
50
|
|
Lw
|
302.6
|
ng/µl
|
1.52
|
DNA
|
50
|
|
Lw
|
263.2
|
ng/µl
|
1.58
|
DNA
|
50
|
|
Lw(Reblank)
|
|||||
|
Lr
|
291.3
|
ng/µl
|
1.44
|
DNA
|
50
|
|
Lr
|
288
|
ng/µl
|
1.44
|
DNA
|
50
|
|
Lr
|
117.1
|
ng/µl
|
1.43
|
DNA
|
50
|
|
Lr
|
109.6
|
ng/µl
|
1.44
|
DNA
|
50
|
|
Lr
|
289.8
|
ng/µl
|
1.43
|
DNA
|
50
|
|
Lb(Reblank)
|
|||||
|
Lb
|
324.2
|
ng/µl
|
1.55
|
DNA
|
50
|
|
Lb
|
239.3
|
ng/µl
|
1.55
|
DNA
|
50
|
|
Lb
|
317.8
|
ng/µl
|
1.53
|
DNA
|
50
|
|
Lb
|
339.3
|
ng/µl
|
1.54
|
DNA
|
50
|
|
Lb
|
354.1
|
ng/µl
|
1.55
|
DNA
|
50
|
|
Sample ID
|
Nucleic Acid
|
Unit
|
260/280
|
Sample Type
|
Factor
|
|
MTg(Blank)
|
|||||
|
MTg
|
200.8
|
ng/µl
|
1.68
|
DNA
|
50
|
|
MTg
|
209.8
|
ng/µl
|
1.68
|
DNA
|
50
|
|
MTg
|
203
|
ng/µl
|
1.66
|
DNA
|
50
|
|
MTg
|
203.4
|
ng/µl
|
1.68
|
DNA
|
50
|
|
MTg
|
202.5
|
ng/µl
|
1.68
|
DNA
|
50
|
|
MTg(Reblank)
|
|||||
|
MTw
|
205.2
|
ng/µl
|
1.54
|
DNA
|
50
|
|
MTw
|
206.6
|
ng/µl
|
1.53
|
DNA
|
50
|
|
MTw
|
207.8
|
ng/µl
|
1.54
|
DNA
|
50
|
|
MTw
|
207.2
|
ng/µl
|
1.54
|
DNA
|
50
|
|
MTw
|
255.3
|
ng/µl
|
1.52
|
DNA
|
50
|
|
MTr(Reblank)
|
|||||
|
MTr
|
107.5
|
ng/µl
|
1.51
|
DNA
|
50
|
|
MTr
|
117.5
|
ng/µl
|
1.41
|
DNA
|
50
|
|
MTr
|
116.7
|
ng/µl
|
1.53
|
DNA
|
50
|
|
MTr
|
106.6
|
ng/µl
|
1.59
|
DNA
|
50
|
|
MTr
|
113.4
|
ng/µl
|
1.53
|
DNA
|
50
|
|
MTb(Reblank)
|
|||||
|
MTb
|
127.6
|
ng/µl
|
1.55
|
DNA
|
50
|
|
MTb
|
131.4
|
ng/µl
|
1.56
|
DNA
|
50
|
|
MTb
|
127.6
|
ng/µl
|
1.55
|
DNA
|
50
|
|
MTb
|
131.1
|
ng/µl
|
1.56
|
DNA
|
50
|
|
MTb
|
132.4
|
ng/µl
|
1.55
|
DNA
|
50
|
|
MTb
|
152.1
|
ng/µl
|
1.54
|
DNA
|
50
|
|
Sample ID
|
Nucleic Acid
|
Unit
|
260/280
|
Sample Type
|
Factor
|
|
Cg(Blank)
|
|||||
|
Cg
|
1786.8
|
ng/µl
|
1.5
|
DNA
|
50
|
|
Cg
|
1810.2
|
ng/µl
|
1.45
|
DNA
|
50
|
|
Cg
|
1895.7
|
ng/µl
|
1.43
|
DNA
|
50
|
|
Cg
|
1685
|
ng/µl
|
1.44
|
DNA
|
50
|
|
Cg
|
1778.2
|
ng/µl
|
1.42
|
DNA
|
50
|
|
Cw(Reblank)
|
|||||
|
Cw
|
514
|
ng/µl
|
1.46
|
DNA
|
50
|
|
Cw
|
516.5
|
ng/µl
|
1.46
|
DNA
|
50
|
|
Cw
|
519.9
|
ng/µl
|
1.46
|
DNA
|
50
|
|
Cw
|
515.6
|
ng/µl
|
1.45
|
DNA
|
50
|
|
Cw
|
521.2
|
ng/µl
|
1.44
|
DNA
|
50
|
|
Cr(Reblank)
|
|||||
|
Cr
|
309.9
|
ng/µl
|
1.57
|
DNA
|
50
|
|
Cr
|
327.8
|
ng/µl
|
1.57
|
DNA
|
50
|
|
Cr
|
316.4
|
ng/µl
|
1.57
|
DNA
|
50
|
|
Cr
|
313.2
|
ng/µl
|
1.57
|
DNA
|
50
|
|
Cr
|
319.6
|
ng/µl
|
1.56
|
DNA
|
50
|
|
Cb(Reblank)
|
|||||
|
Cb
|
357.1
|
ng/µl
|
1.55
|
DNA
|
50
|
|
Cb
|
345
|
ng/µl
|
1.56
|
DNA
|
50
|
|
Cb
|
345.2
|
ng/µl
|
1.56
|
DNA
|
50
|
|
Cb
|
344.2
|
ng/µl
|
1.56
|
DNA
|
50
|
|
Cb
|
348.2
|
ng/µl
|
1.56
|
DNA
|
50
|
Next
week I’ll work on obtaining averages and plotting the data on graphs which will
perhaps make it easier to see what’s happening but from what I can tell the TE
Boiling protocols significantly yielded more DNA than any other protocol
competing against it. Which is impressive because it’s one of the less
elaborate methods to follow.
There
are a number of variables that I would like to mention but I’ll do that next
week. Until next time, Luis.
