Using AuPreP Gold cDNA synthesis Kit , RNA is reverse transcribed into single-stranded cDNA. The reverse transcriptase (RT) enzyme synthesizes the new cDNA strand at a site determined by the type of primer used: Oligo(dT) primer, random primer or a sequence-specific primer. The First Strand cDNA can then be used as a template for PCR. The reaction mix solution supplied, contains buffer, RT with reduced Rnase H activity, dNTP’s mix with 99% purity & Rnase inhibitor
……sufficient for 50 Rxns. Protocol to Step 1:
http://www.atzlabs.com
NB- The product of the first strand cDNA synthesis can be used directly in PCR. The volume of first strand cDNA synthesis reaction mixture should not comprise more than 1/10 of the total PCR reaction volume. Normally, 2 μl of the first strand cDNA synthesis reaction mixture is used as template for subsequent PCR in 50 μl total volume.
Step 2
Amplification Step-
AuPreP Diamond 2x Double Dye Master Mix is a high density 2X Master Mix containing ultrapure and highly sensitive Taq DNA polymerase, dNTPs, PCR enhancer & stabilizer, buffer with the magnesium concentration optimal for routine PCR, gel loading & tracking dyes.
……sufficient for 50 Rxns.
Protocol to Step 2:
http://www.atzlabs.com
NB - DETAILED PROTOCOLS COVERING PRODUCTS FOR STEP 1 & STEP 2 ARE ALSO PROVIDED SEPARATELY*. PLEASE GO THROUGH THEM WHILE USING THE PRODUCT . (* Web links provided as well)