- In illustration, target = Target DNA
- In illustration, numbers are time when the sample was put in since first samples were put in
2012/08/09
<img src="http://openwetware.org/images/thumb/e/e3/20120809_%E2%91%A1.jpg/800px-20120809_%E2%91%A1.jpg" width="620px">
condition
- We use 10% and 20 % acrylamide gel in erectrophoresis
- We stained the 10% gel with midori green, but the gel wasn't stained clearly(the picture is not here).
- The above picture is the 20% gel stained with sybr gold
result
- Each Selector seemed to hybridize with the target
2012/08/17
<img src="http://openwetware.org/images/thumb/e/e3/20120817_%E2%91%A1.jpg/800px-20120817_%E2%91%A1.jpg" width="620px">
condition
- In order to let the target move between Selectors, we waited 50 minutes before the next step
result
- The materials diffused and the bands weren't clear because we forgot to put 15μl buffer containg 1.25 % concentration of Mg2+ to samples
2012/08/20
<img src="http://openwetware.org/images/thumb/5/5d/20120820_10-_%E2%91%A1.jpg/800px-20120820_10-_%E2%91%A1.jpg" width="620px">
<img src="http://openwetware.org/images/thumb/a/af/201120820_20-_%E2%91%A1.jpg/767px-201120820_20-_%E2%91%A1.jpg" width="620px">
condition
- To let the target move between Selectors, we waited one hour before the next step today. We did electrophoresis at 50V in a refrigerate( at 4 degrees)
- We stain the 10% gel with sybr gold
- We do erectrophoresis in refrigerate （4℃） and constant voltage of 50v
- The bands in the 20% gel were warped and we couldn't get the sufficient result
- The following statement is about the electrophoresis in the 10% gel
result
- As a result, the target hybridized with Selector 2 in the lane of the target, Selector 1, and 2. The target also hybridized with Selector 3 in the lane of the target, Selector 1, 2, and 3
- From this result, Target DNA may be able to move from Selector 1 to Selecor 2, and from Selector 2 to Selector 3
- In the lane of the target, Selector 1 and 2, the band of the target and Selector1 and the band of the target and Selector 2 are both clear. So we have some doubts to say the target actually moved from Selector 1 to Selector 2
- In the lane of the target, Selector 1, 2, and 3, no band is around the place of single-stranded Selector 1. So it's probable we forgot to put it
- In the lane of the target, Selector 1, 2, and 3, no band is around the place of single-stranded Selecto r1. So it's probable we forgot to put Selector 1
- We don't make sure the target moved from Selector 1 to Selector 2. But we can probably say the target moved from Selector 2 to Selector 3
2012/8/21
<img src="http://openwetware.org/images/thumb/b/be/20120821_%E2%91%A1.jpg/800px-20120821_%E2%91%A1.jpg" width="620px">
condition
- We did electrophoresis with the same condition as yesterday, except that we didn't prepare the lane of the target, Selector1, 2, and 3
result
- As a result, in the lane of the target, Selector 1, and 2, the band of Selector 1 cannot be seen. And the band of Selector 2 is seen at the same place as when it was single-stranded
- The target didn't seem to attach either to Selector 1 or 2
- So the Selector 1 and 2 aggregated with themselves or each other
- As the 20% gel is difficult to get the result and it takes a long time to, We'll use only the 10% gel from now on
2012/08/22
<img src="http://openwetware.org/images/thumb/e/e3/20120822_%E2%91%A1.jpg/800px-20120822_%E2%91%A1.jpg" width="620px">
condition
- the voltage of electrophoresis is 50v by constant voltage
- We used 1X TAE added to Mg2+ as a buffer
- The room temperature is 28℃
- We did the electrophoresis for 3 hours
- We used SYBR Gold as a stain for 10 minutes
result
- There were bands of sample mixed target and Selector 1, and target and Selector 2, hybridized each other
- In sample mixed Selector 1 and Selector 3, there were both bands Selector 1 only and Selector 3 only so Selector 1 and Selector 3 don't interact each other
- In sample mixed Selector 2 and Selector 3, We could see a black band at one place
- Maybe Selector 2's hairpin structure and Selector 3's interacted each other.
2012/08/24
<img src="http://openwetware.org/images/thumb/a/af/20120824_%E2%91%A1.jpg/791px-20120824_%E2%91%A1.jpg" width="620px">
condition
- the voltage of electrophoresis is 50v by constant voltage
- We used 1X TAE was added to Mg2+ as a buffer
- The room temperature is 28℃
- We did the electrophoresis for 3 hours
- We used SYBR Gold as a stain for 10 minutes
result
- Selector 1 seem to hybridised with itself
2012/08/28
<img src="http://openwetware.org/images/thumb/5/5a/20120828_%E2%91%A1.jpg/800px-20120828_%E2%91%A1.jpg" width="620px">
condition
- the voltage of electrophoresis is cv 50v
- We used 1X TAE was added to Mg2+ as a buffer
- The room temperature is 33℃
- We did the electrophoresis for 3 hours
- We used SYBR Gold as a stain for 10 minutes
result
- We used new base sequence DNA and Selector 1 didn't hybridize with target, but other results were what we want
- Tm of Selector 1 is lower than today's room temperature, so didn't hybridize
- Tomorrow we will try in the refrigerator
2012/08/29
<img src="http://openwetware.org/images/thumb/4/4f/20120829_%E2%91%A1.jpg/800px-20120829_%E2%91%A1.jpg" width="620px">
condition
- We used 1X TAE was added to Mg2+ as a buffer
- The room temperature is 33℃(4℃ in the refrigerator)
- We did the electrophoresis for 3.5 hours
- We used SYBR Gold as a stain for 10 minutes
result
- Selector 1 didn't hybridize with target. Other samples worked well
- We will try again with cool buffer
2012/08/30
<img src="http://openwetware.org/images/thumb/c/c7/20120830_%E2%91%A1.jpg/800px-20120830_%E2%91%A1.jpg" width="620px">
condition
- We used 1X TAE was added to Mg2+ as a buffer
- The room temperature is 33℃(4℃ in the refrigerator)
- We did the electrophoresis for 3.5 hours in the refrigerator
- We used SYBR Gold as a stain for 10 minutes
result
- The result was what we want
- Swlwctor 1 hybridized with target
