Paulsson:Electroporation competent MC1061


 * Electroporation protocol

Batch 11/27/06, labeled with one red dot, made by Per 1.5 x 10(9) cells per ug (transformation efficiency tested with ts-Rep pDNA) 50ul aliquots, some tubes labeled "2x" contain 100ul of cells FINISHED

Batch 01/16/07, labeled with red ink, made by Per 6 x 10(9) cells per ug (transformation efficiency tested with pCR2.1 pDNA) 95ul aliquots, can use for trfs.FINISHED

Batch 03/09/07, labeled "MC1061" with blue ink, 104ul aliquots, made by Per 1 x 10(9) cells per ug (transformation eff. tested w/ 1ng pSilver's Venus, 50ul cells)FINISHED

Batch 06/22/07, labeled "MC" with blue ink, made by Per 1.4 x 10(9) cells per ug (transformation efficiency tested with 100ul cells and 1ng pCR2.1 pDNA) 100 and 200ul aliquotsFINISHED

Batch 07/07, labeled "MC1061" with black ink, ~200ul aliquots, made by Per use 100 ul per transformationFINISHED

Batch ~03/08, labeled "MC1061" with red ink, ~200ul aliquotsFINISHED use ~95 ul per transformation; transformation efficiency ~1.6 x 10(8) cfu per ug plasmid DNA  made by Anna Andersson (using 500ml conical tubes for spins), tested by Per using pCR2.1 (1ng/ul stock)

Batch 06/25/08, labeled "MC1061" with black ink, 105ul aliquots, made by PerFINISHED 3.4 x 10(8) ampR cfu per ug pDNA (transformation efficiency tested with 50ul cells and 1ng pCR2.1 pDNA)

Parallel batch prepared using 500ml Corning Centrifuge Tubes (rather than std bottle)FINISHED labeled "MC1061" with black ink and one BLUE DOT, 105ul aliquots 3.0 x 10(8) ampR cfu per ug pDNA (transformation efficiency tested with 50ul cells and 1ng pCR2.1 pDNA; ampR cfu per cfu = 1/2000)

Batch 08/15/08, labeled "MC1061" with blue ink, 105ul aliquots (37x from 2L OD 0.5), made by PerFINISHED Prepared using 500ml Corning Centrifuge Tubes 6.0 x 10(8) ampR cfu per ug pDNA (transformation efficiency tested with 50ul cells and 1ng pCR2.1 pDNA)

Batch 10/18/08, in "Competent Cells II" box, labeled "X" with green ink, ~105ul aliquots, made by PerFINISHED Prepared using 500ml Corning Centrifuge Tubes and slower spins than previously, ~65 aliquots from 3L 8.9 x 10(8) ampR cfu per ug pDNA (transformation efficiency tested with 50ul cells and 1ng pCR2.1 pDNA)

Batch 04/11/09, in "Competent Cells II" box, labeled with a black line on cap, ~105ul aliquots, made by PerFINISHED Prepared using 500ml Corning Centrifuge Tubes and slower spins than previously, ~60 aliquots from 3L 3 x 10(9) ampR cfu per ug pDNA (transformation efficiency tested with 30ul cells and 1ng pCR2.1 pDNA in 1mm cuvette) 1.4 x 10(9) ampR cfu per ug pDNA (transformation efficiency tested with 60ul cells and 1ng pCR2.1 pDNA in 2mm cuvette, both grought to 1ml in SOC after transformation and 100ul plated)

Batch 10/01/09, in "Competent Cells II" box, labeled "MC" in red ink, ~150ul aliquots, made by PerFINISHED Prepared using 500ml Corning Centrifuge Tubes, final resuspension in larger volume than expected, ~36 aliquots from 1.5L ~1.6 x 10(9) ampR cfu per ug pDNA (transformation efficiency tested with 72.5ul cells and 1ng pCR2.1 pDNA in 2mm cuvette, brought to 1ml in SOC after transformation, grown 60min at 37°C, 100ul plated on LB/Amp)

Batch 02/11/10, in "Competent Cells II" box, labeled "MC" in red ink, ~155ul aliquots, made by PerFINISHED Prepared using 500ml Corning Centrifuge Tubes, final resuspension in larger volume than expected, ~34 aliquots from 1.5L 1.2 x 10(9) ampR cfu per ug pDNA (transformation efficiency tested with 50ul cells and 1ng pCR2.1 pDNA in 1mm cuvette) 0.8 x 10(9) ampR cfu per ug pDNA (transformation efficiency tested with 77ul cells and 1ng pCR2.1 pDNA in 2mm cuvette, both brought to 1ml in SOC after transformation, grown 60min at 37°C, and 100ul plated on LB/Amp)

Batch 11/12/10, in "Paulsson Lab, Competent Cells" box, tubes labeled "MC" in black ink, 155ul aliquots, made by Per FINISHED Prepared using 500ml Corning Centrifuge Tubes, 72 aliquots from 3L 9 x 10(8) ampR cfu per ug pDNA (transformation efficiency tested with 40ul cells and 1ng pCR2.1 pDNA in 1mm cuvette)

Batch 04/15/11, tubes labeled "MC" in blue ink, 160ul aliquots, in "Paulsson Lab, Competent Cells" box <Br> 5 x 10(8) ampR cfu per ug pDNA (transformation efficiency tested with 50ul cells and 1ng pCR2.1 pDNA in 1mm cuvette)<Br> 1.5 x 10(8) ampR cfu per ug pDNA (transformation efficiency tested with 100ul cells and 2ng pCR2.1 pDNA in 2mm cuvette)<Br> 2 x 10(7) ampR cfu per ug pDNA (transformation efficiency tested with 50ul cells and 1ng pCR2.1 pDNA in 1mm cuvette)<Br> 1 x 10(7) ampR cfu per ug pDNA (transformation efficiency tested with 100ul cells and 2ng pCR2.1 pDNA in 2mm cuvette)<Br>
 * Also made 1:8 diluted batch labeled "MC 1:8" in blue ink, 160ul aliquots, in "Paulsson Lab, Competent Cells II" box <Br>