17/09/13

From 2013.igem.org

(Difference between revisions)
(→Results from previous day)
(→Digestion of new fusion PCR Tod genes)
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==Digestion of new fusion PCR Tod genes==
==Digestion of new fusion PCR Tod genes==
*Digestion with EcorI-HF and PstI-HF using Cutsmart buffer
*Digestion with EcorI-HF and PstI-HF using Cutsmart buffer
 +
*500ng of DNA was digested from the following Tod DNA concentrations:
 +
{|border=1
 +
|Sample||Concentration ng/ul||260/280||260/230
 +
|-
 +
|Tod X||49.5||1.89||1.99
 +
|-
 +
|Tod F||33.9||1.98||1.68
 +
|-
 +
|Tob B||37.4||1.82||1.70
 +
|}
 +
*Volumes for double digest:
{|border=1
{|border=1
|Sample||DNA||Buffer||Water||EcorI-HF||PstI-HF
|Sample||DNA||Buffer||Water||EcorI-HF||PstI-HF

Revision as of 10:23, 18 September 2013

Contents

Results from previous day

  • Background control plate had growth, which was not expected. This suggests that pSB1C3 backbone religated.
  • It was also thought that the Tod genes used were not from the PCR fusion experiment.
  • Same experiment done again, with a new set of Tod genes from a PCR fusion.
  • For digestion of backbone, already digested backbone from 01/08/2013 was used, which was digested with EcoRI and PstI.

Incubation of bacteria from the plates of previous day for mini prep

  • 12 single colonies were picked from each plate and put into 15ml of broth
  • 36 tubes were left incubating overnight at 37C

Sending pGEM-T vector with Tod insert clones to sequencing by PNACL

Digestion of new fusion PCR Tod genes

  • Digestion with EcorI-HF and PstI-HF using Cutsmart buffer
  • 500ng of DNA was digested from the following Tod DNA concentrations:
SampleConcentration ng/ul260/280260/230
Tod X49.51.891.99
Tod F33.91.981.68
Tob B37.41.821.70
  • Volumes for double digest:
SampleDNABufferWaterEcorI-HFPstI-HF
Tod X10.1ul3ul15.90.50.5
Tod F13.4ul3ul12.60.50.5
Tob B14.7ul3ul11.30.50.5
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