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 | |