To repeat the PCR that we did [[Team:Newcastle/25_August_2010|yesterday]] using the correct ''rocF'' primers.
-
-
===Materials and Protocol===
-
-
Please refer to [[Team:Newcastle/PCR|PCR]].
-
-
-
==PCR Purification==
-
-
===Aim===
-
-
To remove unwanted primers, taq polymerase, buffer and salts to obtain pure DNA.
-
-
===Materials and Protocol===
-
-
Please refer to [[Team:Newcastle/PCR_purification|PCR purification]].
-
-
-
==Digestion==
-
-
===Aim===
-
-
To digest the PCR products of pSB1C3 and ''yneA'' from PCR purification.
-
-
===Materials and Protocol===
-
-
Please refer to [[Team:Newcastle/Restriction_digests|restriction digest]].
-
-
===Results, Discussion and Conclusion===
-
-
We run the digested products with gel electrophoresis to determine whether the digest worked.
-
-
==Gel extraction==
-
-
===Aim===
-
-
To purify the DNA of ''yneA'' and pSB1C3 by extracting the bands from the gel after running gel electrophoresis. Concentration of DNA is then checked with NanoDrop.
First transformation of 'Bacillius subtilis 168' with Prrnb-GFP containing YneA
Starch plating
Aims
Yesterday we replica plated Bacillus subtilis 168, that were transformed on Wednesday, onto starch plates.
Today we aim to see whether or not the transformed Bacillus subtilis can breakdown starch. If they cannot break down starch there will be no halo when we test with iodine.
Materials
Starch plates
Pipette tips
Iodine
Results
Unfortunately our colonies had halos. Although the insert has integrated, the colonies have antibiotic restistance, the insert has not integrated at the amyE locus.
"
