Horseshoe Crab Extraction Troubleshooting

Checking Extraction Protocol Again to Increase Yield

Following extraction protocol exactly except there was no PBS wash and the volume of proteinase K was increased to 20ul. All samples were digested for 2 hours.

Images from during extraction

Sample 797 tissue piece (minced):

Sample 797 inside the 300ul DNA/RNA Shield tube:

Sample 821 tissue piece (minced):

Sample 821 inside the 300ul DNA/RNA Shield tube:

Sample 824 tissue piece (minced):

Sample 824 inside the 300ul DNA/RNA Shield tube:

Sample 825 tissue piece (minced):

Sample 825 inside the 300ul DNA/RNA Shield tube:

Sample 797 after 2 hours of incubation:

Sample 821 after 2 hours of incubation:

Sample 824 after 2 hours of incubation:

Sample 825 after 2 hours of incubation:

Broad Range Qubit

reading	standard 1 (RFU)	standard 2 (RFU)	797 (ng/ul)	821 (ng/ul)	824 (ng/ul)	825 (ng/ul)
first	189	21137	35.8	53.4	20.8	26.8
second	-	-	36	53.6	20.6	26.4
average	-	-	35.9	53.5	20.7	26.6

1% Agarose Gel

*This was also run with 8 of the previous samples that Natalie has extracted herself using the same protocol

Written on September 17, 2020