PROJECT 6. Homology Arm Amplification

PROJECT 4. Homology Arms Amplification

PROJECTS 3 and 5. Homology Arm Amplification

Lane code	Amplification	Amplicon size	Project
ND 22_1 ND 22_2	5’ Homology arm amplification	746 bp	Project 3 Sax-2_N_S
ND 22_3 ND 22_4	3’ Homology arm amplification	741 bp
DP_315’F DP_315’R	5’ Homology arm amplification	No result	Project 5 Unc-11_C_S
DP_313’F DP_315’F	3’ Homology arm amplification	No result
AH 21_1 AH 21_2	5’ Homology arm amplification	721 bp
AH 21_3 AH 21_4	3’ Homology arm amplification	795 bp

1.5% Agarose gel was used.

PROJECT 2. Homology Arm Amplification

                     

                 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.

5' homolgy arm result for Project 2