SBB11Ntbk-SuhaniVora

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]