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

From ZhangLabWiki
Jump to navigation Jump to search
>Hosuki78
No edit summary
>Hosuki78
No edit summary
 
(7 intermediate revisions by the same user not shown)
Line 4: Line 4:
*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}}
| align="center" style="background:#f0f0f0;"|'''Chamber #'''
| align="center" |10ng
| align="center" style="background:#f0f0f0;"|'''Volume (uL)'''
||
| align="center" style="background:#f0f0f0;"|'''Mass (pg/uL)'''
| align="center" |10ng
| align="center" style="background:#f0f0f0;"|'''Total (pg)'''
|
| align="center" |1ng
||
| align="center" |1ng
|
| align="center" |100pg
||
| align="center" |100pg
|
| align="center" |10pg
||
| align="center" |10pg
|-
|-
|1 ||5.05 | || | ||  
| align="center" |NTC
|-
||
|2 ||5.42 | || | ||
| align="center" |#2
|-
|
|3 ||5.36 | || | ||
| align="center" |#5
|-
||
|4 ||5.08 | || | ||
| align="center" |#11
|-
|
|5 ||6.87 | || | ||
| align="center" |#13
|-
||
|6 ||5.46 | || | ||
| align="center" |#18
|-
|
|7 ||5.42 | || | ||
| align="center" |#20
|-
||
|8 ||5.24 | || | ||
| align="center" |#22
|-
|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 | || | ||
|-
|20 ||5.40 | || | ||
|-
|21 ||6.02 | || | ||  
|-
|22 ||4.93 | || | ||
|-
|23 ||5.43 | || | ||
|-
|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)