Noi/NOTES/2013-2-27: Difference between revisions
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'''Probe prep''' | '''Probe prep''' | ||
- Waiting for seed oligos: expected to come next Wednesday (March6, 2013) | - Waiting for seed oligos: expected to come next Wednesday (March6, 2013) | ||
- eMIP_CA1_F and eMIP_CA1_R primer for expansion PCR and production PCR (very little left, ordered 2/28/2013) | - eMIP_CA1_F and eMIP_CA1_R primer for expansion PCR and production PCR (very little left, ordered 2/28/2013) ; arrived 3/04/2013 -> resuspend 100uM | ||
- 2x KAPA SYBG FAST qPCR MM | - 2x KAPA SYBG FAST qPCR MM | ||
- Nt.AlwI (NEB) and Nb.BsrDI (NEB) -> ordered 2/26/2013 | - Nt.AlwI (NEB) and Nb.BsrDI (NEB) -> ordered 2/26/2013 ; arrived 3/04/2013 | ||
- Qiaquick columns, TBU gel, Nanosep columns | - Qiaquick columns, TBU gel, Nanosep columns | ||
'''Padlock capture''' | '''Padlock capture''' | ||
Note: There is no gap for this capture, so only need Ampligase | Note: There is no gap for this capture, so only need Ampligase | ||
- Ampligase and buffer | - Ampligase and buffer: yes | ||
- ExoI/ExoIII | - ExoI/ExoIII: yes | ||
- Amplification primers (Matt) | - Amplification primers (Matt) | ||
* Need to check with Dr. Zhang about the gDNA, RNA and cDNA amount in capture. The expected size of citcularized DNA (no gap --> 200bp?) |
Revision as of 21:11, 5 March 2013
Plan for probe preparation for Matt's project
- I will be preparing probes using the same method as I did for LC Sciences oligoes, [[1]]
- Check reagents to make sure I have all reagents ready for probe prep and padlock capture
Probe prep - Waiting for seed oligos: expected to come next Wednesday (March6, 2013) - eMIP_CA1_F and eMIP_CA1_R primer for expansion PCR and production PCR (very little left, ordered 2/28/2013) ; arrived 3/04/2013 -> resuspend 100uM - 2x KAPA SYBG FAST qPCR MM - Nt.AlwI (NEB) and Nb.BsrDI (NEB) -> ordered 2/26/2013 ; arrived 3/04/2013 - Qiaquick columns, TBU gel, Nanosep columns Padlock capture Note: There is no gap for this capture, so only need Ampligase - Ampligase and buffer: yes - ExoI/ExoIII: yes - Amplification primers (Matt)
- Need to check with Dr. Zhang about the gDNA, RNA and cDNA amount in capture. The expected size of citcularized DNA (no gap --> 200bp?)