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Team:The University of Alberta/24 July 2008
From 2008.igem.org
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*Did mini-preps of J61003 because we were running out | *Did mini-preps of J61003 because we were running out | ||
*Digested BisDA/B with single digests using AgeI and NgoMIV to test the enzymes because weve been getting cruddy digests when using both of them | *Digested BisDA/B with single digests using AgeI and NgoMIV to test the enzymes because weve been getting cruddy digests when using both of them | ||
| + | **'''UPDATE''': It would appear that AgeI is cutting but NgoMIV isn't. Could be a buffer issue? | ||
| + | |||
==System 2== | ==System 2== | ||
Today we have taken the ligation from yesterday and transformed it as well as digested it. | Today we have taken the ligation from yesterday and transformed it as well as digested it. | ||
*if all goes well the digested product should be B0034+TETR | *if all goes well the digested product should be B0034+TETR | ||
**Column Purified the digested product. | **Column Purified the digested product. | ||
| - | * | + | **Then ligated the B0034+TETR with LacIQERE giving us a complete system 2. |
| - | *Digested pTet with Spe/Pst. | + | **PCRed the digested product i.e tetR+BOO34 |
| + | *Digested pTet with Spe/Pst. Column purified the digested product. | ||
*Digested RFP with Xba/Pst. | *Digested RFP with Xba/Pst. | ||
==System 3== | ==System 3== | ||
*Digested 5ul of J61003 with Xba/Pst; this will let us insert the Tryp biobrick digested and ligated yesterday | *Digested 5ul of J61003 with Xba/Pst; this will let us insert the Tryp biobrick digested and ligated yesterday | ||
**Ran the digested J6 on a gel; gel purified | **Ran the digested J6 on a gel; gel purified | ||
Latest revision as of 21:20, 24 July 2008
General
- Did mini-preps of J61003 because we were running out
- Digested BisDA/B with single digests using AgeI and NgoMIV to test the enzymes because weve been getting cruddy digests when using both of them
- UPDATE: It would appear that AgeI is cutting but NgoMIV isn't. Could be a buffer issue?
System 2
Today we have taken the ligation from yesterday and transformed it as well as digested it.
- if all goes well the digested product should be B0034+TETR
- Column Purified the digested product.
- Then ligated the B0034+TETR with LacIQERE giving us a complete system 2.
- PCRed the digested product i.e tetR+BOO34
- Digested pTet with Spe/Pst. Column purified the digested product.
- Digested RFP with Xba/Pst.
System 3
- Digested 5ul of J61003 with Xba/Pst; this will let us insert the Tryp biobrick digested and ligated yesterday
- Ran the digested J6 on a gel; gel purified
