QPCR: Difference between revisions
Updated with smaller sample volumes |
m added how to make the 100 uM stock: "-the 100uM stock is prepared by adding 227 uL of distilled water to 22.7nmol of a gene as an example-" |
||
| Line 5: | Line 5: | ||
*SYBR Green PCR Master Mix Applied Biosystems (ThermoFisher Catalog # 4367659; [https://www.thermofisher.com/order/catalog/product/4367659 Vendor Link]) | *SYBR Green PCR Master Mix Applied Biosystems (ThermoFisher Catalog # 4367659; [https://www.thermofisher.com/order/catalog/product/4367659 Vendor Link]) | ||
*384 well qPCR plate (ThermoFisher Catalog # 4309849) and covers (Catalog # 4360954) | *384 well qPCR plate (ThermoFisher Catalog # 4309849) and covers (Catalog # 4360954) | ||
*Primers (Dilute to 0.4 uM mixture of fwd and rev. From 100 uM stocks this is 4uL Forward Primer, 4 uL Reverse Primer and 992 uL Water). This primer mix can be stored at -20 in a Working Primers box. Design primers according to [[Primer Design for qPCR]] | *Primers (Dilute to 0.4 uM mixture of fwd and rev. From 100 uM stocks- the 100uM stock is prepared by adding 227 uL of distilled water to 22.7nmol of a gene as an example- this is 4uL Forward Primer, 4 uL Reverse Primer and 992 uL Water). This primer mix can be stored at -20 in a Working Primers box. Design primers according to [[Primer Design for qPCR]] | ||
===Plate Preparation=== | ===Plate Preparation=== | ||