Latest revision as of 08:57, 19 August 2008

Molecular Biology

We try by different ways to assemble biobricks

We(Vincent) did a mistake with the kanamycin stock, we put 1000 time less the dose.

We start a cell culture of all interesting biobricks so we can do a new mini-prep tomorrow, because the mini-prep that we use are not good(kanamycin 1000 time less concentrated).

E1010 and F1610

We tried last week to assemble two big parts because it is easier to purify them. We did the ligation and transformation. The result was great, all of the ten plates worked. Today, we decide to do a mini-prep of these cells to purify the DNA. Then we want to check the plasmid so we digest a small quantity of them with EcoRI and SpeI to prepare it for a gel tomorrow.

by Gel

We try again purifying insert and vector usign a gel.

We try to assemble :

R0040/B0034

R0071/B0034

and I1466/I1433

We have successfully isolated from the gel : R0040, R0071 and B0034

We try changing strategy to isolate I1466 and I14033.

I1466/I14033

I14033 is small. We keep it in the vector. I1466 is big we use it as an insert. We cut with SpeI and PstI. We digest the plasmid and we are going to do the ligation tomorrow.

Without gel, use PCR with biobrick primer

Tomorrow, we plan on using the new mini-preps to amplify DNA by PCR, this way we don't need to use gel purification (which is hard to do with small fragments).

Addition : We still need gel purification after we digest the PCR product, so that enzymes and other fragments are removed. However this way we use less of our mini-prep material.

http://openwetware.org/wiki/Knight:Annealing_and_primer_extension_with_Klenow_polymerase

@@ Line 3: / Line 3: @@
 =Molecular Biology=
-We try by different way to assemble biobrick
+We try by different ways to assemble biobricks
-We(Vincent) do a mistake with kanamycin stock, we put 1000 time less the dose.
+We(Vincent) did a mistake with the kanamycin stock, we put 1000 time less the dose.
-We start cell culture of all interestings biobrick to do tomorrow fresh mini-prep. Because the mini-prep that we use are not good(kanamycin 1000 time less concentrate).
+We start a cell culture of all interesting biobricks so we can do a new mini-prep tomorrow, because the mini-prep that we use are not good(kanamycin 1000 time less concentrated).
 ==E1010 and F1610==
-We try last week to assemble two big parts because it is easy to purify it.
+We tried last week to assemble two big parts because it is easier to purify them.
-We do ligation and transformation last week. The result was great all of the ten plates work.
+We did the ligation and transformation. The result was great, all of the ten plates worked.
-Monday, we decide to do a mini prep of these cells to purify DNA. Then we want to check the plasmid so we digest a small quantity of them with EcoRI and SpeI to prepare it to do a gel tomorrow.
+Today, we decide to do a mini-prep of these cells to purify the DNA. Then we want to check the plasmid so we digest a small quantity of them with EcoRI and SpeI to prepare it for a gel tomorrow.
 ==by Gel==
-We try again to purify insert and vector by Gel.
+We try again purifying insert and vector usign a gel.
 We try to assemble :
@@ Line 25: / Line 25: @@
 and I1466/I1433
-We successful have isolate from Gel : R0040, R0071 and B0034
+We have successfully isolated from the gel : R0040, R0071 and B0034
-We try to change strategy of isolate I1466 and I14033
+We try changing strategy to isolate I1466 and I14033.
 ==I1466/I14033==
-I14033 is small. We let it in the vector. I1466 is big we use it to a insert.
+I14033 is small. We keep it in the vector. I1466 is big we use it as an insert.
 We cut with SpeI and PstI.
-We digest the plasmid and we are going to do tomorrow ligation.
+We digest the plasmid and we are going to do the ligation tomorrow.
 ==Without gel, use PCR with biobrick primer==
-We will start tomorrow with new mini-prep to amplify DNA by PCR, by this way we don't need to use Gel purification(that is hard with small fragment).
+Tomorrow, we plan on using the new mini-preps to amplify DNA by PCR, this way we don't need to use gel purification (which is hard to do with small fragments).
+Addition : We still need gel purification after we digest the PCR product, so that enzymes and other fragments are removed. However this way we use less of our mini-prep material.
 http://openwetware.org/wiki/Knight:Annealing_and_primer_extension_with_Klenow_polymerase

EPF-Lausanne/11 August 2008

From 2008.igem.org