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xRDL: 1st Year After Christmas - Coggle Diagram
xRDL: 1st Year After Christmas
Reading: 3 papers per day
mRNA Display
RaPID
FIT
PPI and Drug Discovery
RiPPs
Writing
Write your progress every 2 weeks in a report style -> can be in onenote
Experiments
T4 Rnl
Time course experiment to reduce the ligation time
Cause of streaking -> plan experiments to reduce the streaking -> maybe ask Sam
RNAse inhibitor experiment -> order
do a no enzyme control in experiments
PhoRtcB
PEG optimisation
Oligo LC-MS
RNA Clean up did not seem to work -> use proteinase K treatment to remove the protein before adding to the column
or high speed centrifuge in 4 C after heating at 70 C for 10 min.
EcoRtcB
Order from NEB
Clone out the enzyme to remove the unnecessary tags in the sequence
Quantifying the Gel bands
research online how to do this with ImageJ
Done this but I don't think it is the most useful for the project aims -> we can still look at the image and assess the intensity by eye since the method is semi quantitative at best and to really quantify we need to add a standard in the sample that does not change so that we can normalise the densitometry results based on that -> this can be done but it is not worth the effort just yet since the literature does not report a definitive way to do this
RaPID
Thursday with Issy from LW group
Repeats
repeat of the variable PEG8000 experiment but with the optimised ligation time
do a nanodrop of the purified proteins ?
RNA clean up kit?
add proteinase K to the sample prior to clean up kit
the rna recovery is quite good but there still is some sort of RNA loss to the protein
Destaining ?
do two gels of the same experiment one destained and one not
stain for 15 min next time
Coursework
6-Month Report
Course
Extra-Cirrucular
Volunteering
Philosophy
Economy