ON TESTIS AND EPlDlDYMlS OF RATS - Pondicherry University ...

22.91 pg of 4-androstcne 3,17-dione was dissolved in 100 mL of distilled
water.
1.9 17p-Hydroxysteroid dehydrogenase
1.9.1 Sodium pyrophosphate buffer (1 00 pM; pH 9.0)
13.3 rng of tetrasodium pyrophosphate was dissolved and made up to 500 mL
with distilled water and pH was adjusted to 9.0.
1.9.2 NADPH (0.5 mM)
2.08 mg of NADPH was dissolved in 5 mL of distilled water
2.86 mg of androstene 3,17-dione was dissolved in I0 mL of 50% ethanol
1.10 Estimation of deoryribonucleic acids (DNA)
1.10.1 Perchloric acid (PCA)
Perchloric acid 16.66 mL (60%) was made upto 100 mL with distilled water.
1.10.2 Diphenylamine reagent
1.5 g of diphenylamine reagent was dissolved in 100 rnL of glacial acetic acid.
To this 1.5 mL of concentrated sulphuric acid was added and stored at 4°C in
amber coloured bottle.
To every 20 mL of this reagent, 0.1 mL of 1.6% aqueous acetaldehyde was
added to potentiate the colour development.
1.10.3 DNA standard
2 mg of calf thymus DNA was dissolved in 10 mL of hot 5% trichloroacetic
acid to give a concentration of 200 pg/ mL and stored at 4°C.
1.1 1 Estimation of ribonudeic acid (RNA)
1.1 1 .I Orcinol reagent
I g of ferric chloride was dissolved in 1 L of concentrated hydrochloric acid
and 35 mL of 6% orcinol was added.