Kun:LabNotes/ExonomeSeq/2007-12-31: Difference between revisions

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(New page: ==Colony PCR== Perform PCR on 16 colonies from each of the three plates: E55kv1(200+350), E55kv1(250+450), CpGPCR. 2X Taq master mix: 15ul 10uM M13 primer mix: 1ul H2O...)
 
 
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94C 3min -> (94C 30sec -> 54C 30sec -> 72C 45sec) x 35 -> 72C 3min -> 4C hold.
94C 3min -> (94C 30sec -> 54C 30sec -> 72C 45sec) x 35 -> 72C 3min -> 4C hold.
Sequencing results:
{| {{table}}
| align="center" style="background:#f0f0f0;"|''''''
| align="center" style="background:#f0f0f0;"|'''PCR amplicon'''
| align="center" style="background:#f0f0f0;"|'''Linker seq'''
| align="center" style="background:#f0f0f0;"|'''H1'''
| align="center" style="background:#f0f0f0;"|'''H2'''
| align="center" style="background:#f0f0f0;"|'''Insert'''
| align="center" style="background:#f0f0f0;"|'''Note:'''
|-
| 1B-M13R||180bp||correct||chr19:41778717||Chr19:49995389||AP1&AP2||It seems to come from a chimeric amplicon during the amplification of agilent library
|-
| 1C-M13R|| || || || || ||Vector seq only
|-
| 3B-M13R||235bp||correct|| || ||AP1&AP2||It seems to come from a chimeric amplicon during the amplification of agilent library
|-
| 3E-M13R|||| || |||| ||Vector seq only
|-
| 3G-M13R||400bp||correct||correct||correct||248bp, NM_011047605-9.4||
|-
| 3H-M13R||250bp||||||||||Self-ligation of probe
|-
| 4B-M13R||397bp||correct||correct||correct||247bp, NM_001008496-7.2||
|-
|
|}

Latest revision as of 21:02, 3 January 2008

Colony PCR[edit]

Perform PCR on 16 colonies from each of the three plates: E55kv1(200+350), E55kv1(250+450), CpGPCR.

    2X Taq master mix:    15ul
    10uM M13 primer mix:   1ul
    H2O:                  14ul

94C 3min -> (94C 30sec -> 54C 30sec -> 72C 45sec) x 35 -> 72C 3min -> 4C hold.

Sequencing results:

' PCR amplicon Linker seq H1 H2 Insert Note:
1B-M13R 180bp correct chr19:41778717 Chr19:49995389 AP1&AP2 It seems to come from a chimeric amplicon during the amplification of agilent library
1C-M13R Vector seq only
3B-M13R 235bp correct AP1&AP2 It seems to come from a chimeric amplicon during the amplification of agilent library
3E-M13R Vector seq only
3G-M13R 400bp correct correct correct 248bp, NM_011047605-9.4
3H-M13R 250bp Self-ligation of probe
4B-M13R 397bp correct correct correct 247bp, NM_001008496-7.2