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Team:Johns Hopkins/Notebook/GROUP 3: Short two way stops
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of BBa_K110012 was correct. That clone was cut and ligated into iGEM vector. A PCR will be tried again for 11-->13, however direct | of BBa_K110012 was correct. That clone was cut and ligated into iGEM vector. A PCR will be tried again for 11-->13, however direct | ||
ligation into iGEM vector will be attempted to evade problems with incomplete products ligating into PGEM-T. | ligation into iGEM vector will be attempted to evade problems with incomplete products ligating into PGEM-T. | ||
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The Two way stop biobrick was then attached to the end of the Fluorescent Protien Venus YFP, using iGEM standard assembly. | The Two way stop biobrick was then attached to the end of the Fluorescent Protien Venus YFP, using iGEM standard assembly. | ||
This Venus YFP had come from iGEM headquarters, as we as still waiting for sequence verification of our biobricks. | This Venus YFP had come from iGEM headquarters, as we as still waiting for sequence verification of our biobricks. | ||
Revision as of 13:38, 25 August 2008
GROUP 3: Short two way stops
Date: August 25, 2008 Status report by: James Part no.: BBa_K110011 -> BBa_K110013 Part Description: Short Two way Stops Summary: Sequence files from parts sent for sequencing showed incorrect sequences for 3/4 biobricks sent in. However 1 clone of BBa_K110012 was correct. That clone was cut and ligated into iGEM vector. A PCR will be tried again for 11-->13, however direct ligation into iGEM vector will be attempted to evade problems with incomplete products ligating into PGEM-T. The Two way stop biobrick was then attached to the end of the Fluorescent Protien Venus YFP, using iGEM standard assembly. This Venus YFP had come from iGEM headquarters, as we as still waiting for sequence verification of our biobricks. We have yet to run the product on the gel, but the expected band length should be around 900bp. (700bp Fluorescent Protein + 200bp Two way stop)
Date 8/12/08 Status report by: Raghav and James Part no.:BBa_K110011 -> BBa_K110013; BBa_K110010;17 Part Description: Summary:Short two way stops should be sent out for sequencing this week Needed so much DNA to visualize one band of short two way stop. Sapphire FP progress has slowed, due to extraneous bands in gel and extraneous sequence from the trace files. transformations from the biobrick book were tried again with very poor results... 1 COLONY... POSSIBLY THE BIOBRICK BOOK IS UPSETTING.
Date 8/5/08 Status report by: James Part no.: BBa_K110011 -> BBa_K110013; BBa_K110010;17 Part Description: Short two way stops; and Sapphire Fluorescent Proteins Summary: The sequence data from last week, turned out all right both orientations of Sapphire FP should be correct sequences Sequence Data HOORAY! Last weeks miniprep and digestion yielded no DNA, so this week the minprep was repeated and the digestion of the prodeuct can be found here RE digest [http://www.jhu.edu/iGEM/Group3:ShortTwo-wayStops/2008-8-5.Two%20way%20stop%20digestion%208/05/08.Jasper;%20James;%20Raghav.html Two way stop digestion 8/05/08]
Date 7/29/08 Status report by: James Part no.: BBa_K110011 -> BBa_K110013; BBa_K110010;17 Part Description: Short two way stops Summary: Sequencing of BBa_K110010;17 (Sapphire FP) revealed confusing, if not bad sequences.Sequence Data PCR cloning and transformation was completed sucessfully last week on the Short two way stops A Miniprep was done and a RE digest will be completed soon (possibly tonight)
Date: 7/22/2008 Status report by: James Part no.: BBa_K110011 -> BBa_K110013 Part Description: Short two way stops Summary: RE digestion showed no insert the plasmid of our clones, thus the PCR was done again. [http://www.jhu.edu/iGEM/Group3:ShortTwo-wayStops/2008-7-23.Short%20way%20stops.James.html Short way stops]
Date: 7/21/08 status report by: James Part no.: BBa_K110011 Part Description: Between-bud 27-W FRS2-C LtR Part Location (in build a genome lab): In James and Jasper's PCR product Box, Stainless Steel 4 degree PCR successful?; Yes Cloning of PCR product successful: Yes [http://www.jhu.edu/iGEM/Group3:ShortTwo-wayStops/2008-7-22.Short%202Way%20Stop,%20Alpha%20Promoters,%20&%20Sapphire%20FP.Allison%20Suarez%20and%20Nate%20Sotuyo%20.html Short 2Way Stop, Alpha Promoters, & Sapphire FP] Sequencing of cloned PCR product successful: No Joining of validated part to adjacent part(s) status: Not done Problems to be solved: Current status of this part: Miniprep digestion yielded no product, PCR; ligation; transformation; and miniprep will be repeated by this Wednesday (07/23/08)
status report by: James Part no.: BBa_K110012 Part Description: Between STE2-W and BST1-C LtR Part Location (in build a genome lab): In James and Jasper's PCR product Box, Stainless Steel 4 degree PCR successful?; Yes Cloning of PCR product successful: Yes [http://www.jhu.edu/iGEM/Group3:ShortTwo-wayStops/2008-7-22.Short%202Way%20Stop,%20Alpha%20Promoters,%20&%20Sapphire%20FP.Allison%20Suarez%20and%20Nate%20Sotuyo%20.html Short 2Way Stop, Alpha Promoters, & Sapphire FP] Sequencing of cloned PCR product successful: No Joining of validated part to adjacent part(s) status: Not done Problems to be solved: Current status of this part: Miniprep digestion yielded no product, PCR; ligation; transformation; and miniprep will be repeated by this Wednesday (07/23/08)
status report by: James Part no.: BBa_K110013 Part Description: Between-SWP82-W and EMP47-C LtR Part Location (in build a genome lab): In James and Jasper's PCR product Box, Stainless Steel 4 degree PCR successful?; No Cloning of PCR product successful: No Sequencing of cloned PCR product successful: No Joining of validated part to adjacent part(s) status: Not done Problems to be solved: The PCR of this part yielded a very large product Current status of this part:
