SBB11Ntbk-SuhaniVora: Difference between revisions
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Place in thermocycler at 37 deg for 1 hr.<br> | Place in thermocycler at 37 deg for 1 hr.<br> | ||
Lane 3: sbb1121 (P_rfaQ) [1031 bp] | |||
Lane 4: sbb1134 (P_malK) [525 bp] |
Revision as of 11:55, 22 February 2011
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Suhani Vora 14:08, 15 February 2011 (EST)
Today we set up PCR reactions for our promoter parts.
-Resuspended oligos PmalK_R and SV_06 to 100 um.
-Diluted primers to 10 um.
-Set up PCR for cloning using Expand Polymerase
1. P_malk
2. P_nlpA
3. P_rfaQ
Thermocycler Program: 2K55
Suhani Vora 13:24, 17 February 2011 (EST)
Ran PCR samples of P_malK, P_nlpA, P_rfaQ on analytical gel.
2ul Sample + 5 uL Loading Dye Buffer
Sample 1: P_malK (525 bp)
Sample 2: P_nlpA (542 bp)
Sample 3: P_rfaQ (1031 bp)
Cleaned with Zymo Cleanup : May have used wrong buffer
All PCR products for the class were thrown out, re-done by Prof. Anderson.
Suhani Vora 13:37, 22 February 2011 (EST)
P_nlpA PCR failed yesterday. Will have to move on without that part.
EcoRI/BamHI Digest of P_rfaQ (sbb1121) and P_malK (sbb1134)
Place in thermocycler at 37 deg for 1 hr.
Lane 3: sbb1121 (P_rfaQ) [1031 bp] Lane 4: sbb1134 (P_malK) [525 bp]