Four to six milliliters of blood was collected into EDTA-coated tubes from toddlers on visits when they had no fever, cold, flu, infections or other illnesses, or use of medications for illnesses 72 hours prior blood draw. Temperature was also taken at the time of blood draw. Blood samples were passed over a LeukoLOCK filter (Ambion, Austin, TX, USA) to capture and stabilize leukocytes and immediately placed in a -20°C freezer. Total RNA was extracted following standard procedures and manufacturer’s instructions (Ambion, Austin, TX, USA). LeukoLOCK disks (Ambion Cat #1933) were freed from RNA-later and Tri-reagent (Ambion Cat #9738) was used to flush out the captured lymphocyte and lyse the cells. RNA was subsequently precipitated with ethanol and purified through washing and cartridge-based steps. The quality of mRNA samples was quantified by the RNA Integrity Number (RIN), values of 7.0 or greater were considered acceptable [45 (link)], and all processed RNA samples passed RIN quality control. Quantification of RNA was performed using Nanodrop (Thermo Scientific, Wilmington, DE, USA). Samples were prepped in 96-well plates at the concentration of 25 ng/μl.
Leukolock disks
Manufactured by Thermo Fisher Scientific
LeukoLOCK disks are a laboratory product designed to capture and isolate leukocytes (white blood cells) from biological samples. The disks utilize a proprietary capture technology to selectively bind and retain leukocytes while allowing other cellular components to pass through. This process enables the extraction and concentration of leukocytes for further analysis or downstream applications.
Automatically generated - may contain errors
4 protocols using leukolock disks
1
RNA Extraction from Toddler Blood
2
Toddler Blood Leukocyte Extraction
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Four to six milliliters of blood was collected into EDTA-coated tubes from toddlers on visits when they had no fever, cold, flu, infections or other illnesses, or use of medications for illnesses 72 hours prior to blood draw. Temperature was also taken at the time of blood draw. Blood samples were passed over a LeukoLOCK filter (Ambion, Austin, TX, USA) to capture and stabilize leukocytes and immediately placed in a −20 °C freezer. Total RNA was extracted following standard procedures and manufacturer’s instructions (Ambion, Austin, TX, USA). LeukoLOCK disks (Ambion, Cat #1933) were freed from RNA-later and Tri-reagent (Ambion, Cat #9738) was used to flush out the captured lymphocyte and lyse the cells. RNA was subsequently precipitated with ethanol and purified through washing and cartridge-based steps. The quality of messenger RNA samples was quantified by the RNA integrity number (RIN), with values of 7.0 or greater considered acceptable [45 (link)], and all processed RNA samples passed RIN quality control. Quantification of RNA was performed using Nanodrop (Thermo Scientific, Wilmington, DE, USA). Samples were prepped in 96-well plates at the concentration of 25 ng/µl.
3
Toddler Blood Leukocyte RNA Extraction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Four to six milliliters of blood was collected into EDTA-coated tubes from toddlers on visits when they had no fever, cold, flu, infections or other illnesses, or use of medications for illnesses 72 hours prior blood draw. Blood samples were passed over a LeukoLOCK™ filter (Ambion, Austin, TX, USA) to capture and stabilize leukocytes and immediately placed in a 20°C freezer. Total RNA was extracted following standard procedures and manufacturer’s instructions (Ambion, Austin, TX, USA). LeukoLOCK disks (Ambion Cat #1933) were freed from RNA-later and Tri-reagent (Ambion Cat #9738) was used to flush out the captured lymphocyte and lyse the cells. RNA was subsequently precipitated with ethanol and purified though washing and cartridge-based steps. The quality of mRNA samples was quantified by the RNA Integrity Number (RIN), values of 7.0 or greater were considered acceptable59 (link), and all processed RNA samples passed RIN quality control. Quantification of RNA was performed using Nanodrop (Thermo Scientific, Wilmington, DE, USA). Samples were prep in 96-well plates at the concentration of 25 ng/µl.
4
Toddler Blood Leukocyte RNA Extraction
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