Magnetic mRNA Isolation Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- If the lysate viscous than normal. Pass these cell or tissue lysates several times through a 21-gauge needle. If the yield of mRNA is low Increase the ratio of beads to Lysis/Binding Buffer or add RNase-free Proteinase K to the sample lysate directly before the 10 minute binding incubation

Protocol tips
- If the lysate viscous than normal. Pass these cell or tissue lysates several times through a 21-gauge needle. If the yield of mRNA is low Increase the ratio of beads to Lysis/Binding Buffer or add RNase-free Proteinase K to the sample lysate directly before the 10 minute binding incubation

Publication protocol

RNA from the rat tissues was extracted using the Magnetic mRNA Isolation kit (No. S1550S; New England Biolabs, Inc.). cDNA was synthesized using 200 ng mRNA, random hexamer primers, and RevertAid Premium (Thermo Fisher Scientific). PCR amplification was performed on 0.2 ng single-stranded cDNA with 2U Thermus aquaticus (Taq) DNA polymerase (Axon). Cycling conditions were 94°C for 3 min, 94°C for 30 s, 60°C for 20 s, 72°C for 30 s for 32 cycles, respectively, and 72°C for 8 min. Controls were performed on the cloned olfactory and retinal ANO 2 splice variants (see “Molecular cloning of rat ANO 1 and ANO 2”). The bands were resolved by gel electrophoresis and verified by sequencing.

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Manufacturer protocol

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