Hosuk:LabNotesMDA/2013-6-21: Difference between revisions

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*MDA run [[Hosuk:LabNotesMDA/2013-6-19|@06/19]]
*MDA run [[Hosuk:LabNotesMDA/2013-6-19|@06/19]]
*qPCR result was not correct because of using wrong primers --> re-run
*qPCR result was not correct because of using wrong primers --> re-run
**For the initial test, 7 samples - 2, 5, 8, 11, 13, 18, 20, 22 were run.
**For the initial test, 7 samples - 2, 5, 11, 13, 18, 20, 22 were run.
 
 
=====Samples=====
 
{| {{table}}
{| {{table}}
|Chamber #
| align="center" |10ng
||
|Volume (uL)
||
|Mass. Est. (pg/uL)
||
|Total (pg)
|-
|1
||5.05
|
||
|
||
|-
|2
||5.42
|
||
|
||
|-
|3
||5.36
|
||
|
||
|-
|4
||5.08
|
||
|
||
|-
|5
||6.87
|
||
|
||
|-
|6
||5.46
|
||
|
||
|-
|7
||5.42
|
||
|
||
|-
|8
||5.24
|
||
|
||
|-
|9
||5.22
|
||
|
||
|-
|10
||4.80
|
||
|
||
|-
|11
||5.34
|
||
|
||
|-
|12
||5.15
|
||
|
||
|-
|13
||4.25
|
||
|
||
|-
|14
||5.01
|
||
|
||
|-
|15
||4.83
|
||
|
||
|-
|16
||4.58
|
||
|
||
|-
|17
||5.93
|
||
|
||
|-
|18
||5.74
|
||
|
||
|-
|19
||5.19
|
||
||
| align="center" |10ng
|
|
| align="center" |1ng
||
||
|-
| align="center" |1ng
|20
||5.40
|
|
| align="center" |100pg
||
||
| align="center" |100pg
|
|
| align="center" |10pg
||
||
| align="center" |10pg
|-
|-
|21
| align="center" |NTC
||6.02
|
||
|
||
|-
|22
||4.93
|
||
||
| align="center" |#2
|
|
| align="center" |#5
||
||
|-
| align="center" |#11
|23
||5.43
|
|
| align="center" |#13
||
||
| align="center" |#18
|
|
| align="center" |#20
||
||
| align="center" |#22
|-
|-
|24
||4.60
|
||
|
||
|
|
|}
|}
====Result --> Fail!====
*Ladders were correct, but samples were not shown any PCR amplification.
*MDA didn't work.
*Possible reason would be
**Experimental error, or
**Re-hybridization while Neutralization?
*Run one more time with the same condition but keeping 1 min. incubation carefully while neutralization, (not too long)

Latest revision as of 18:30, 21 June 2013

MDA with PGP1F, 4th - qPCR[edit]

  • MDA run @06/19
  • qPCR result was not correct because of using wrong primers --> re-run
    • For the initial test, 7 samples - 2, 5, 11, 13, 18, 20, 22 were run.
10ng 10ng 1ng 1ng 100pg 100pg 10pg 10pg
NTC #2 #5 #11 #13 #18 #20 #22


Result --> Fail![edit]

  • Ladders were correct, but samples were not shown any PCR amplification.
  • MDA didn't work.
  • Possible reason would be
    • Experimental error, or
    • Re-hybridization while Neutralization?
  • Run one more time with the same condition but keeping 1 min. incubation carefully while neutralization, (not too long)