Sunday, July 31

From 2011.igem.org

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Once again tried tranformation into competent cells.  Used 2 protocols 1st protocol from New England Biodabs High Efficiency Protocol for C2987 cells
Once again tried tranformation into competent cells.  Used 2 protocols 1st protocol from New England Biodabs High Efficiency Protocol for C2987 cells
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Added 1 ul of each of material from tubes 1, 3, 2B, 2, 1B, (dot or scribble),4A,  5A, 5B to 50 ul of competent cells (was not enough competent cells to do all tube samples   
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Added 1 ul of each of material from tubes 1, 3, 2B, 2, 1B, (dot or scribble),4A,  5A, 5B to 50 ul of competent cells (was not enough competent cells to do all tube samples)  
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Added control PNC19 to one of the competent cell vials, and put no dna into the second competent cell vial.
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Added control PUC19 to one of the competent cell vials, and put no dna into the second competent cell vial.
  Left on ice for 30 minutes.  heat shocked tubes 2 at a time for 45 secs.  Placed on ice for 5 minutes.  be
  Left on ice for 30 minutes.  heat shocked tubes 2 at a time for 45 secs.  Placed on ice for 5 minutes.  be
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Pipetted 950 ul of SOC into each tube
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Pipetted 950 ul of room temp SOC into each tube
put in warm room shaker, shook for 1 hour
put in warm room shaker, shook for 1 hour
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Used a Protocol from Tom, from Chung, Niemela, and Miller  "One-step preparation of competent Escherichia coli:  Transformation and storage of bacterial cells in the same solution," Proc Natl. Acad. Sci Vol 86, pp 2172-2175, April 1989 {{LabReportBmore}}
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Also Used a Protocol from Tom, from Chung, Niemela, and Miller  "One-step preparation of competent Escherichia coli:  Transformation and storage of bacterial cells in the same solution," Proc Natl. Acad. Sci Vol 86, pp 2172-2175, April 1989
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Added 1ul of each material from tubes 3B and 4A to 900 uL of 2xTSS.  No dna and PUC19  were use as controls
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put into warm room, and shook for 1 hour
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Plated all (both protocols) using hockey stick dipped in 80% ethanol, to smear 100 uL of each sample and at 17:30 plates were put into warm room
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Excess material was put in freezer, labelled "C2987 Transformation" or "TSS Transformation" with today's date, in green rack
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Total hours 5.5 SS
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 +
{{LabReportBmore}}

Revision as of 21:57, 31 July 2011

Once again tried tranformation into competent cells. Used 2 protocols 1st protocol from New England Biodabs High Efficiency Protocol for C2987 cells Added 1 ul of each of material from tubes 1, 3, 2B, 2, 1B, (dot or scribble),4A, 5A, 5B to 50 ul of competent cells (was not enough competent cells to do all tube samples) Added control PUC19 to one of the competent cell vials, and put no dna into the second competent cell vial.

Left on ice for 30 minutes.  heat shocked tubes 2 at a time for 45 secs.  Placed on ice for 5 minutes.  be

Pipetted 950 ul of room temp SOC into each tube put in warm room shaker, shook for 1 hour

Also Used a Protocol from Tom, from Chung, Niemela, and Miller "One-step preparation of competent Escherichia coli: Transformation and storage of bacterial cells in the same solution," Proc Natl. Acad. Sci Vol 86, pp 2172-2175, April 1989 Added 1ul of each material from tubes 3B and 4A to 900 uL of 2xTSS. No dna and PUC19 were use as controls put into warm room, and shook for 1 hour

Plated all (both protocols) using hockey stick dipped in 80% ethanol, to smear 100 uL of each sample and at 17:30 plates were put into warm room

Excess material was put in freezer, labelled "C2987 Transformation" or "TSS Transformation" with today's date, in green rack

Total hours 5.5 SS

Team:Baltimore/Notebook

Hours