DNase treated the 07:12 set of Abalone Dg RNA. Also, respec’d the RNA prior to proceeding. Followed Ambion’s Turbo DNA-free Kit rigorous protocol. Here are the calcs/workup for the DNase treatment. This used previously untreated RNA.
Tag Archives: DNase
DNase Treatment – Abalone Dg DNased RNA 20090610
This is the 4th time this RNA has been DNased. Performed using Ambion’s Tubro DNA-free Kit. Followed the “rigorous” treatment protocol.
DNase Treatment – MV hemocyte RNA from yesterday
Samples 3326: B23, A25, A22, B14, A21, A10 B22 came up positive for gDNA still. These were retreated according to Ambion protocol with a brand new Turbo DNA-free DNase kit. Additionally, I tested all three existing kits by “spiking” 19uL of H2O with 1uL (~200ng) of gigas gDNA; one tube for each kit and an untreated sample. Will qPCR to see if gDNA removal was successful.
DNase Treatment – MV hemocyte RNA from earlier today
The entire 20uL of RNA were treated with Ambion’s Turbo DNA-free kit according to protocol and spec’d.
Mac performed qPCR on the DNase-treated RNAs to verify removal of residual gDNA.
Results: Samples 3326: B23, A25, A22, B14, A21, A10 B22 came up positive for residual gDNA. Will retreat these samples and qPCR again.
DNase Treatment – Abalone Dg DNased RNA from 20090605
This is the 3rd time this will be performed on these samples. Hopefully using a different (i.e. new) kit will work. Not sure what’s going on here. Performed according to protocol.
DNase Treatment – Abalone Dg DNased RNA from 20090605
Had to re-treat these samples due to residual gDNA that was NOT eliminated the first time through. Possible old Ambion kit??
DNase Treatment – Abalone Dg RNA isolated yesterday and from 20090518
RNA from yesterday was treated according to Ambion Turbo DNA-free protocol to remove gDNA contamination. Work up is here.
DNase Treatment – Abalone Dg RNA isolated yesterday
RNA from yesterday was treated according to Ambion Turbo DNA-free protocol to remove gDNA contamination. Work up is here. Will check for residual gDNA contamination once I’ve finished with another set of RNA isolations from the remainder of Abalone Dg tissue.
DNase Treatment (Rigorous!) – Mac’s gigas RNA/Re-DNased RNA from 20090507 & 20090508, respectively
Followed the rigorous protocol for Ambion’s Turbo DNA-free protocol for the following RNAs:
BB#11-20
DH#11-20
Used 10ug of RNA (200ng/uL) in 50uL as directed. Here are the calcs for FF and DH.
Followed standard protocol on DNased samples from 20090508:
BB#1-10
DH#1-10
The standard protocol should be fine for these samples, since the procedure worked on Friday for some of them.
DNase Treatment – Oyster RNA from yesterday
Yesterday’s qPCR indicated that all of the RNA still contained gDNA contamination. So, took 10ug of RNA from BB #1-10 and DH#1-10 (calcs/workup BB and DH) and brought the volumes up to 50uL with 0.1% DEPC-H2O. Processed the samples according to Ambion Tubrbo DNA-free protocol. Will proceed with qPCR immediately.
