AlanFung:LabNotes/CTCF/2010-3-24: Difference between revisions

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>Alan6017518
>Alan6017518
Line 5: Line 5:
[[File:ZhangLab_2 2010-03-24 11hr 35min.jpg]]
[[File:ZhangLab_2 2010-03-24 11hr 35min.jpg]]
* I think both the nanaodrop and gel quantification are biased, for the nanaodrop it gives out an extremely high amount of concentration, this usually happens because nanaodrop cannot differentiate between any contamination. As for the gel image the band at 100bp is much stronger than the ladder band which hits that the concentration is higher than 18.95ng/ul. In this case I would go with the qubit quantification.
* I think both the nanaodrop and gel quantification are biased, for the nanaodrop it gives out an extremely high amount of concentration, this usually happens because nanaodrop cannot differentiate between any contamination. As for the gel image the band at 100bp is much stronger than the ladder band which hits that the concentration is higher than 18.95ng/ul. In this case I would go with the qubit quantification.
==Library Prep with new Jurkat PCR product==
*Concentration:48.4ng/ul
*BP:100BP
*setup: 100ng,50ng, 25ng, 10ng, 5ng and -ve control
==End Repair==
{| {{table}}
| ||100ng||50ng||25ng||10ng||5ng||-ve
|-
| DNA||2.07||1.03||5.17||2.07||1.03||0
|-
| End Repair Buffer||10||10||10||10||10||10
|-
| End Repair Enzyme mix||5||5||5||5||5||5
|-
| ddH2o||82.93||83.97||79.83||82.93||83.97||85
|-
| Total Volume||100||100||100||100||100||100
|-
|
|}

Revision as of 21:06, 24 March 2010

Quantification of Jurkat PCR Product

Nanodrop: 250.7ng/ul
Qubit: 48.4ng/ul
Gel Quant: 18.95ng/ul

File:ZhangLab 2 2010-03-24 11hr 35min.jpg

  • I think both the nanaodrop and gel quantification are biased, for the nanaodrop it gives out an extremely high amount of concentration, this usually happens because nanaodrop cannot differentiate between any contamination. As for the gel image the band at 100bp is much stronger than the ladder band which hits that the concentration is higher than 18.95ng/ul. In this case I would go with the qubit quantification.

Library Prep with new Jurkat PCR product

  • Concentration:48.4ng/ul
  • BP:100BP
  • setup: 100ng,50ng, 25ng, 10ng, 5ng and -ve control

End Repair

100ng 50ng 25ng 10ng 5ng -ve
DNA 2.07 1.03 5.17 2.07 1.03 0
End Repair Buffer 10 10 10 10 10 10
End Repair Enzyme mix 5 5 5 5 5 5
ddH2o 82.93 83.97 79.83 82.93 83.97 85
Total Volume 100 100 100 100 100 100