Hosuk:LabNotes/2013-4-23: Difference between revisions
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**Protocol | **Protocol | ||
**[[File:RCAReationProtocolinTube.jpg|350px]] | **[[File:RCAReationProtocolinTube.jpg|350px]] | ||
*Mixing Cy3-Adapter | *Mixing Cy3-Adapter | ||
Line 16: | Line 18: | ||
**S2 : -CircLigase II + Cy3-Adapter | **S2 : -CircLigase II + Cy3-Adapter | ||
**S3 : H2O + Cy3-Adapter | **S3 : H2O + Cy3-Adapter | ||
=====Result===== | |||
*According to the image of RCA with +CircLigase II, there are RCA product. | |||
*So, CircLigase II reaction was done. | |||
*Again, circularized 50bp DNA ran faster than linear in the 20% TBU gel. | |||
{| {{table}} | |||
| S1:RCA, +CircLigase | |||
|| | |||
| S2:RCA, -CircLigase | |||
|| | |||
| S3:H2O | |||
|| | |||
|- | |||
|([[Media:CircII_Custom_RCA_Cy3Adapter_20x_fig02.tif|Original]]) | |||
|| | |||
|([[Media:-CircII(noenzyme)_Custom_RCA_Cy3Adapter_20x_fig02.tif|Original]]) | |||
|| | |||
|([[Media:H2O_Cy3Adapter_20x_fig02.tif|Original]]) | |||
|- | |||
|[[File:CircII_Custom_RCA_Cy3Adapter_20x_fig02.jpg|300px]] | |||
|| | |||
|[[File:-CircII(noenzyme)_Custom_RCA_Cy3Adapter_20x_fig02.jpg|300px]] | |||
|| | |||
|[[File:H2O_Cy3Adapter_20x_fig02.jpg|300px]] | |||
|} |
Revision as of 00:27, 24 April 2013
RCA test with the product of CircLigase II reaction
Goal
- Verify that there are circularized DNA in the CircLigase II reaction sample
Experiment Procedure
- Run RCA
- S1 : CircLigase II reaction sample (18hr. @ 30°C)
- S2 : -CircLigase II sample (No CircLigase II Enzyme while CircLigase II reaction) (18hr. @ 30°C)
- Protocol
- File:RCAReationProtocolinTube.jpg
- Mixing Cy3-Adapter
- 4.5uL of each RCA sample + 0.5uL of 100uM Cy3-Adapter
- S1 : +CircLigase II + Cy3-Adapter
- S2 : -CircLigase II + Cy3-Adapter
- S3 : H2O + Cy3-Adapter
Result
- According to the image of RCA with +CircLigase II, there are RCA product.
- So, CircLigase II reaction was done.
- Again, circularized 50bp DNA ran faster than linear in the 20% TBU gel.
S1:RCA, +CircLigase | S2:RCA, -CircLigase | S3:H2O | |||
(Original) | (Original) | (Original) | |||
File:CircII Custom RCA Cy3Adapter 20x fig02.jpg | File:-CircII(noenzyme) Custom RCA Cy3Adapter 20x fig02.jpg | File:H2O Cy3Adapter 20x fig02.jpg |