Homology arm Result for Project 2 (Gel image 2 after 10 mins extra run time)
In this step of project 2 (N-terminal knock-in of unc-11 long isoform), we amplified our homology arms from C.elegans genomic DNA.
Lane 1 and 2: 5' homology arm (ISOLATE CODES?)
Lane 3 and 4: 3' homology arm (ISOLATE CODES?)
Lane 5: 1kb ladder from NEB
Lane 6 and 7: 5' homology arm (ISOLATE CODES?)
Lane 8 and 9: 3' homology arm (ISOLATE CODES?)
The predicted sizes of each arm are 699bp for 5' and 605bp for 3'. Pairs of primers used:
5' Homology- Forward primer:TGTAAAACGACGGCCAGTCGGGCAAATCCCTGCGGAAATGAGA
5' Homology- Reverse primer: TCCTCTCCCTTGGAGACCATgtcgagcagcaaggggggt
3' Homology- Forward primer: TGATCAGCGAGGAAGACTTGATGcagactatcgagaaagc
3' Homology- Reverse primer: AACAGCTATGACCATGTTATtcgagagctaatttcagcga
Comparing the sizes of the bands with 1kb ladder, the bands run from 600pb to 700bp as what expected.
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ReplyDeleteIn this step of project 2 (unc-11, N terminal fusion, long isoform), we tried to build the map of homology arms selected from unc-11 gene of C.elegans that will be working in the pDD286 gene.
ReplyDeleteOn the gel:
Lane 1 and 2: 5' homology arm (TN's samples)
Lane 3 and 4: 3' homology arm (TN's samples)
Lane 5: 1kb ladder from NEB
Lane 6 and 7: 5' homology arm (RV's samples)
Lane 8 and 9: 3' homology arm (RV's sample)
The predicted sizes of each arm are 699bp for 5' and 605bp for 3'. Pairs of primers used:
5' Homology- Forward primer:TGTAAAACGACGGCCAGTCGGGCAAATCCCTGCGGAAATGAGA
5' Homology- Reverse primer: TCCTCTCCCTTGGAGACCATgtcgagcagcaaggggggt
3' Homology- Forward primer: TGATCAGCGAGGAAGACTTGATGcagactatcgagaaagc
3' Homology- Reverse primer: AACAGCTATGACCATGTTATtcgagagctaatttcagcga
Comparing the sizes of the bands with 1kb ladder, the bands run from 600pb to 700bp as what expected.
Lane 6-9 tube codes RV 21-1, RV 21-2, RV-3 and RV 21-4
ReplyDelete