User:Anthony Salvagno/Notebook/Research/2009/03/17/D&L Day 4
Today, so far and to do:
- P/C extraction of overnight digestion
- EtOH precipitation
- Make minigel
- Run minigel comparing digestion with uncut
- If digestion worked well, we will do the ligation.
Gel Results and Remarks
|Tube Number||Vol plasmid DNA||Vol gDNA||Vol Buffer 10x||Vol T4 DNA Ligase||Vol H2O||Total Vol|
- Allow to incubate at 16C overnight (in PCR chamber thing).
This time the ratios, by weight, are (V:I, vector:insert):
- V only
Digesting Another Try
|Tube Number||Vol gDNA||Vol and Type 10x Buffer||Vol BSA 10x||Vol XhoI||Vol H2O||Total Vol|
- Add 3.5uL more XhoI after ~2 hours
Mary Ann told Kelly and I we might get better digestion if we use less DNA, so we lowered the amount in the sample.